Information on EC 3.1.31.1 - micrococcal nuclease

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY
3.1.31.1
-
RECOMMENDED NAME
GeneOntology No.
micrococcal nuclease
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
endonucleolytic cleavage to nucleoside 3'-phosphates and 3'-phosphooligonucleotide end-products
show the reaction diagram
mechanism
-
endonucleolytic cleavage to nucleoside 3'-phosphates and 3'-phosphooligonucleotide end-products
show the reaction diagram
mechanism
Staphylococcus aureus Foggi, Staphylococcus aureus V8
-
-
endonucleolytic cleavage to nucleoside 3'-phosphates and 3'-phosphooligonucleotide end-products
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of phosphoric ester
-
-
-
-
hydrolysis of phosphoric ester
-
-
hydrolysis of phosphoric ester
-
-
hydrolysis of phosphoric ester
-
-
hydrolysis of phosphoric ester
Staphylococcus aureus KCCM 11335
-
-
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
AtTSN1
Q9FLT0
-
AtTSN2
Q8VZG7
-
EC 3.1.4.7
-
-
formerly
-
Epstein-Barr virusencoded transcription factor 2 co-activator p100
Q7KZF4
-
HsTSN
Q7KZF4
-
micrococcal DNase
-
-
-
-
micrococcal endonuclease
-
-
-
-
Micrococcal nuclease
-
-
-
-
Micrococcal nuclease
-
-
MN
-
-
-
-
NUC1
-
gene name
NUC1
Staphylococcus aureus RN4220
-
gene name
-
nuclease 8V
-
-
-
-
nuclease T
-
-
-
-
nuclease T'
-
-
-
-
nuclease, micrococcal
-
-
-
-
nuclease, staphylococcal
-
-
-
-
P100
Q7KZF4
-
PaTSN
Q0JRI3
-
ribonucleate (deoxyribo-nucleate) 3'-nucleotidohydrolase
-
-
-
-
ribonucleate (deoxyribonucleate) 3'-nucleotidohydrolase
-
-
-
-
S. aureus nuclease
-
-
-
-
SNA
Staphylococcus aureus KCCM 11335
-
-
-
snake venom phosphodiesterase
-
-
-
-
SNase
-
-
-
-
SNAseR
-
analogue of SNAse A EC 3.1.4.7, in which hexapeptide is appended to the alanine of SNAse A
SND1
Q7KZF4
-
spleen endonuclease
-
-
-
-
spleen phosphodiesterase
-
-
-
-
staph nuclease
-
-
-
-
staphylococcal nuclease
-
-
-
-
staphylococcal nuclease
Q7KZF4
-
staphylococcal nuclease
Staphylococcus aureus RN4220
-
-
-
Staphylococcal nuclease A
-
-
Staphylococcal nuclease A
Staphylococcus aureus KCCM 11335
-
-
-
staphylococcal nuclease domain-containing protein 1
Q7KZF4
-
staphylococcus aureus nuclease
-
-
-
-
staphylococcus aureus nuclease
-
-
staphylococcus aureus nuclease
-
Nuc, named NucB when secreted as an active 168-amino acid-long polypeptide
staphylococcus aureus nuclease
Staphylococcus aureus USA 300 LAC
-
-
-
staphylococcus aureus nuclease B
-
-
-
-
Staphylococcus aureus nuclease homologue
-
-
thermonuclease
-
-
-
-
thermonuclease
-
-
TNase
-
-
-
-
TSN
Q7KZF4
-
TSN
Q0JRI3
-
tudor staphylococcal nuclease
Q8VZG7, Q9FLT0
-
tudor staphylococcal nuclease
Q7KZF4
-
tudor staphylococcal nuclease
Q0JRI3
-
tudor staphylococcal nuclease
-
-
CAS REGISTRY NUMBER
COMMENTARY
9013-53-0
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
Escherichia coli pFOG
pFOG
-
-
Manually annotated by BRENDA team
complementation of the Lactococcus lactis secretion machinery with Bacillus subtilis SecDF improves secretion of staphylococcal nuclease
-
-
Manually annotated by BRENDA team
strains 3D7 and W2mef, enzyme expressed in all three blood stages: ring, trophozoite and schizont stage
-
-
Manually annotated by BRENDA team
a community-acquired CA-MRSA strain, gene nuc
-
-
Manually annotated by BRENDA team
ATCC 6538P
-
-
Manually annotated by BRENDA team
mutant D21E
-
-
Manually annotated by BRENDA team
mutant P117G
-
-
Manually annotated by BRENDA team
mutant V66W
-
-
Manually annotated by BRENDA team
mutants L25A L36A 61G62 I72A 90P91 Y91A I92A Y93A 94A95 L103G 103A104 G107V L108A L125A L125G; strain Foggi Worthington
-
-
Manually annotated by BRENDA team
native state protein and engineered proteins showing a disordered state under physiological conditions
-
-
Manually annotated by BRENDA team
strain Foggi
-
-
Manually annotated by BRENDA team
strain Foggi Worthington; strain V8
-
-
Manually annotated by BRENDA team
strain KCCM 11335, Korean Culture Center of Microorganisms, Korea
-
-
Manually annotated by BRENDA team
unfolded enzyme
-
-
Manually annotated by BRENDA team
unfolded enzyme
Uniprot
Manually annotated by BRENDA team
var.aureus
-
-
Manually annotated by BRENDA team
Staphylococcus aureus 209P
209P
-
-
Manually annotated by BRENDA team
Staphylococcus aureus Foggi
strain Foggi
-
-
Manually annotated by BRENDA team
Staphylococcus aureus Foggi
strain Foggi Worthington
-
-
Manually annotated by BRENDA team
Staphylococcus aureus KCCM 11335
strain KCCM 11335, Korean Culture Center of Microorganisms, Korea
-
-
Manually annotated by BRENDA team
Staphylococcus aureus RN4220
gene nuc1
-
-
Manually annotated by BRENDA team
Staphylococcus aureus USA 300 LAC
a community-acquired CA-MRSA strain, gene nuc
-
-
Manually annotated by BRENDA team
Staphylococcus aureus V8
strain V8
-
-
Manually annotated by BRENDA team
gene nuc
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
malfunction
-
when the nuc1 gene is knocked out, the ability of Staphylococcus aureus strains to form a biofilm significantly increased
malfunction
Staphylococcus aureus RN4220
-
when the nuc1 gene is knocked out, the ability of Staphylococcus aureus strains to form a biofilm significantly increased
-
metabolism
-
programmed cell death pathways, Tudor staphylococcal nuclease is a new component of the human programmed degradome and is cleaved by caspase-3 between the Tudor and SN5 domains
physiological function
-
staphylococcal nuclease domain-containing protein 1 is a component of the RNA-induced splicing complex that mediates RNA interference, leading to degradation of specific mRNAs
physiological function
-
Staphylococcal nuclease degrades both DNA and RNA
physiological function
-
ribonuclease activity, control of gene expression by activating transcription, subsequent mRNA splicing, regulation of RNA silencing and in the pathway that edits and destroys double-stranded RNA, uncleavable Tudor staphylococcal nuclease stimulates cell proliferation and protects cells from death
physiological function
-
Staphylococcus aureus nuclease mediates resistance against entrapment and killing within neutrophil extracellular traps, which consist of a nuclear DNA backbone associated with antimicrobial peptides, histones and proteases that provide a matrix to entrap and kill various microbes. Promoting role of Staphylococcus aureus nuclease in neutrophil extracellular traps degradation and virulence in a murine respiratory tract infection model, overview. Staphylococcus aureus nuclease facilitates evasion from neutrophil extracellular trap entrapment, and mediates resistance against extracellular killing by neutrophils
physiological function
-
presence and ionization of Lys66, buried in the hydrophobic core of a stabilized variant of staphylococcal nuclease, affect conformation and dynamics. The neutral Lys66 affects slow conformational fluctuations globally, whereas the effects of the charged form are localized to the region immediately surrounding position 66, when Lys66 is charged the protein expands, structural reorganization triggered by ionization of the internal Lys66, detailed overview
physiological function
-
staphylococcal nuclease is an important virulence factor of Staphylococcus aureus. Biofilm development can be prevented in staphylococcal nuclease-producing strains of Staphylococcus aureus, direct relationship between staphylococcal nuclease production and the prevention of biofilm development, overview. Staphylococcal nuclease affects biofilm formation by other bacteria, such as Pseudomonas aeruginosa and Staphylococcus epidermis. Only bacterial strains that do not possess the nuc1 gene are able to form biofilms
physiological function
Staphylococcus aureus RN4220
-
staphylococcal nuclease is an important virulence factor of Staphylococcus aureus. Biofilm development can be prevented in staphylococcal nuclease-producing strains of Staphylococcus aureus, direct relationship between staphylococcal nuclease production and the prevention of biofilm development, overview. Staphylococcal nuclease affects biofilm formation by other bacteria, such as Pseudomonas aeruginosa and Staphylococcus epidermis. Only bacterial strains that do not possess the nuc1 gene are able to form biofilms
-
physiological function
Staphylococcus aureus USA 300 LAC
-
Staphylococcus aureus nuclease mediates resistance against entrapment and killing within neutrophil extracellular traps, which consist of a nuclear DNA backbone associated with antimicrobial peptides, histones and proteases that provide a matrix to entrap and kill various microbes. Promoting role of Staphylococcus aureus nuclease in neutrophil extracellular traps degradation and virulence in a murine respiratory tract infection model, overview. Staphylococcus aureus nuclease facilitates evasion from neutrophil extracellular trap entrapment, and mediates resistance against extracellular killing by neutrophils
-
metabolism
Q0JRI3
programmed cell death, enzyme is a part of the stress-induced cell-death degradome during both developmental and stress-induced cell deaths
additional information
-
thermodynamic parameters derived from urea-induced unfolding of mutant SNase140 and mutant SNase141 in comparison with those of wild-type SNase and mutant SNase140 in the presence of calcium, backbone dynamics, detailed overview. Mutant SNase140 unfolds easily compared to mutant SNase141 and wild-type SNase
additional information
-
residue W140 is critical to SNase structure and function
additional information
-
MNase mapping of the enhancer chromatin structure in the Drosophila melanogaster embryo to analyse chromatin structure in a developmental setting, and identification of structural changes on a cis-regulatory element targeted by the Knirps repressor, method development and optimization, overview
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
3'-O-acetylnitrophenyl-pdT + H2O
?
show the reaction diagram
-
-
-
-
?
5'-chloromethyl-pdTp-nitrophenyl + H2O
?
show the reaction diagram
-
-
-
-
?
5'-O-acetyl-dTp-nitrophenyl + H2O
?
show the reaction diagram
-
-
-
-
?
5'-sulfate-dTp-nitrophenyl + H2O
?
show the reaction diagram
-
-
-
-
?
DNA
?
show the reaction diagram
-
-
-
-
?
DNA + H2O
3'-deoxymononucleotides + dinucleotides
show the reaction diagram
-
-
-
-
-
DNA + H2O
3'-deoxymononucleotides + dinucleotides
show the reaction diagram
-
-
-
?
DNA + H2O
3'-deoxymononucleotides + dinucleotides
show the reaction diagram
-
-
-
-
?
DNA + H2O
3'-deoxymononucleotides + dinucleotides
show the reaction diagram
-
-
-
-
?
DNA + H2O
3'-deoxymononucleotides + dinucleotides
show the reaction diagram
-
-
+ oligonucleotides terminated by 3'-phosphate, produced only in incomplete digestion
-
DNA + H2O
3'-deoxymononucleotides + dinucleotides
show the reaction diagram
-
in native DNA Xp-dTp and Xp-dAp bonds are preferentially attached in denaturated DNA random cleavage, denaturated DNA is hydrolyzed more rapidly than native DNA
-
-
?
DNA + H2O
3'-deoxymononucleotides + dinucleotides
show the reaction diagram
Staphylococcus aureus V8
-
-
-
-
?
DNA + H2O
3'-deoxymononucleotides + dinucleotides
show the reaction diagram
Staphylococcus aureus V8
-
in native DNA Xp-dTp and Xp-dAp bonds are preferentially attached in denaturated DNA random cleavage, denaturated DNA is hydrolyzed more rapidly than native DNA
-
-
?
DNA + H2O
3'-deoxymononucleotides + dinucleotides
show the reaction diagram
Staphylococcus aureus Foggi
-
in native DNA Xp-dTp and Xp-dAp bonds are preferentially attached in denaturated DNA random cleavage, denaturated DNA is hydrolyzed more rapidly than native DNA
-
-
?
DNA + H2O
3'-deoxymononucleotides + dinucleotides
show the reaction diagram
Staphylococcus aureus KCCM 11335
-
-
-
-
?
dTp-nitrophenyl + H2O
?
show the reaction diagram
-
-
-
-
?
GFP-ssDNA + H2O
?
show the reaction diagram
-
-
partial degradation
-
?
GFP-ssRNA + H2O
?
show the reaction diagram
-
-
complete degradation
-
?
M13mp18 DNA + H2O
?
show the reaction diagram
-
circular single stranded DNA
-
-
?
nitrophenyl-pdT + H2O
?
show the reaction diagram
Staphylococcus aureus, Staphylococcus aureus V8, Staphylococcus aureus Foggi
-
-
-
-
?
nitrophenyl-pdTp + H2O
?
show the reaction diagram
-
-
-
-
?
nitrophenyl-pdTpdTp-nitrophenyl + H2O
?
show the reaction diagram
-
-
-
-
?
RNA + H2O
nucleoside 3'-phosphates + dinucleotides
show the reaction diagram
-
-
-
-
-
RNA + H2O
nucleoside 3'-phosphates + dinucleotides
show the reaction diagram
-
-
-
-
?
RNA + H2O
nucleoside 3'-phosphates + dinucleotides
show the reaction diagram
-
-
-
?
RNA + H2O
nucleoside 3'-phosphates + dinucleotides
show the reaction diagram
-
-
-
?
RNA + H2O
nucleoside 3'-phosphates + dinucleotides
show the reaction diagram
-
-
-
-
?
RNA + H2O
nucleoside 3'-phosphates + dinucleotides
show the reaction diagram
-
-
dinucleotides terminated by 3'-phosphates
?
RNA + H2O
nucleoside 3'-phosphates + dinucleotides
show the reaction diagram
-
-
nucleoside 3'-phosphates of both purines and pyrimidines
?
RNA + H2O
nucleoside 3'-phosphates + dinucleotides
show the reaction diagram
Staphylococcus aureus V8
-
-
-
-
?
RNA + H2O
nucleoside 3'-phosphates + dinucleotides
show the reaction diagram
Staphylococcus aureus V8, Staphylococcus aureus Foggi
-
-
-
?
ss-DNA + H2O
?
show the reaction diagram
-
single strand salmon sperm DNA, obtained by boiling for 30 min and rapid cooling on ice
-
-
?
ssDNA + H2O
?
show the reaction diagram
-
single stranded salmon sperm DNA
-
-
?
T2 DNA + H2O
?
show the reaction diagram
-
-
-
-
?
methyl-pdTp-nitrophenyl + H2O
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
-
-
-
-
additional information
?
-
-
specificity
-
-
-
additional information
?
-
-
inhibition when a 5'-phosphomonoester end group is present in an oligonucleotide, best substrates oligonucleotides with a 3'-phosphomonoester end group
-
-
-
additional information
?
-
-
substrate masking: binding of RNA by EGTA-inactivated enzyme results in artifactual inhibition of RNA processing
-
-
-
additional information
?
-
-
enzyme does not cleave the 2',3'-cyclic phosphate derivates of the ribonucleosides
-
-
-
additional information
?
-
-
staphylococcal nuclease R, an analogue of the enzyme has the same activity and structural feature as the wild type enzyme, poly-his-nuclease R can be used both for removal of contaminated DNA and RNA and for separating the enzyme from target proteins
-
-
-
additional information
?
-
-
essential enzyme in the life cycle of Plasmodium falciparum, GFP-dsDNA and GFP-dsRNA are not used as substrates
-
-
-
additional information
?
-
-
micrococcal nuclease induces double-strand breaks within nucleosome linker regions, and with more extensive digestion, single-strand nicks within the nucleosome itself
-
-
-
additional information
?
-
-
substrate is single-strand salmon sperm DNA
-
-
-
additional information
?
-
Staphylococcus aureus V8, Staphylococcus aureus Foggi
-
specificity
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
DNA + H2O
3'-deoxymononucleotides + dinucleotides
show the reaction diagram
-
-
-
-
?
DNA + H2O
3'-deoxymononucleotides + dinucleotides
show the reaction diagram
-
-
-
-
?
DNA + H2O
3'-deoxymononucleotides + dinucleotides
show the reaction diagram
Staphylococcus aureus KCCM 11335
-
-
-
-
?
additional information
?
-
-
staphylococcal nuclease R, an analogue of the enzyme has the same activity and structural feature as the wild type enzyme, poly-his-nuclease R can be used both for removal of contaminated DNA and RNA and for separating the enzyme from target proteins
-
-
-
additional information
?
-
-
essential enzyme in the life cycle of Plasmodium falciparum
-
-
-
additional information
?
-
-
micrococcal nuclease induces double-strand breaks within nucleosome linker regions, and with more extensive digestion, single-strand nicks within the nucleosome itself
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
hydrolysis of DNA and RNA is completely dependent on Ca2+, KM for wild-type enzyme is 0.113 mM
Ca2+
-
required
Ca2+
-
1 mM, unfolding forces in the presence of calcium ion are slightly increased
Ca2+
-
enzyme is Ca2+ dependent for its activity
Ca2+
-
activity is Ca2+ dependent, 10 mM are included in assay buffer
Ca2+
-
5 mM CaCl2 are included in assay medium
Ca2+
Q0JRI3
30 mM CaCl2 are included in assay medium
Ca2+
-
the carboxylic groups in the active site of SNase are known to bind Ca2+
Ca2+
-
required
Cu2+
-
minimal activation if Ca2+ is replaced by Cu2+
Fe2+
-
minimal activation if Ca2+ is replaced by Fe2+
Sr2+
-
DNase but no RNase activity if Ca2+ is replaced Sr2+
Mg2+
-
required
additional information
-
examination of salt sensitivity of pka-values (chemical shifts analyzed by NMR while titrating) His8, His46, His121 and His124. Tested variants D21N/T33V/T41I/S59A/P117G/A128A mutant, E75A mutant, E75Q mutant, and E101A mutant in 0.01 M, 0.10 M, and 1.0 M KCl
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2'-deoxythymidine 3',5'-diphosphate
-
at 100 microM concentration of pdTp, parasites show block in development, most of the parasites appear dead or shrunken within six hours after inhibitor treatment
3',5'-deoxythymidine diphosphate
-
addition to the cell culture medium blocks further growth
adenosine 3',5'-diphosphate
-
able to induce folding of mutant proteins into the native state
caspase-3
-
Tudor staphylococcal nuclease, a multifunctional regulator of gene expression, is cleaved by caspase-3 during apoptosis, this cleavage impairs the ability of Tudor staphylococcal nuclease to activate mRNA splicing, inhibits its ribonuclease activity and is important for the execution of apoptosis, cleavage of enzyme lowers its nuclease activity by almost 50%
-
deoxythymidine 3',5'-bisphosphate
-
nuclease activity of enzyme is highly sensitive to deoxythymidine 3',5'-bisphosphate, a specific inhibitor of staphylococcal nuclease, whereas the inactive analogue deoxythymidine 3'-phosphate has no inhibitory effect
deoxythymidine 3',5'-bisphosphate
Q0JRI3
nuclease activity of PaTSN is highly sensitive to deoxythymidine 3',5'-bisphosphate, a specific inhibitor of staphylococcal nuclease, whereas the inactive analogue deoxythymidine 3'-phosphate has no inhibitory effect
deoxythymidine 3',5'-diphosphate
-
in the presence of pdTp, unfolding forces increase drastically
metacaspase mcII-Pa
-
metacaspase mcII-Pa cleaves the phylogenetically conserved protein Tudor staphylococcal nuclease during both developmental and stress-induced programmed cell death
-
metacaspase mcII-Pa
Q0JRI3
metacaspase mcII-Pa processes recombinant enzyme into 5 major fragments, inactivation of mcII-Pa either by the inhibitor EGR-chloromethyl ketone or through mutation of catalytic Cys139 blocked Tudor staphylococcal nuclease fragmentation, proteolysis of endogenous enzyme correlates with metacaspase activity, reaching the maximum level in early embryos and declining to non-detectable levels in mature embryos, cleavage of enzyme by mcII-Pa leads to a 90% decrease in ribonuclease activity, whereas inactivation of mcII-Pa by either EGR-cmk or mutation of catalytic Cys139 left TSN intact and fully active
-
oligonucleotides
-
with 5'-phosphate end group
thymidine 3',5'-bisphosphate
-
competitive
Mononucleotides
-
with 5'-phosphate end group
additional information
-
induction of apoptosis in HCT116 colon carcinoma cells with 5-fluorouracil leads to increased caspase-3-like activity and cleavage of endogenous Tudor staphylococcal nuclease into 2 fragments with the same molecular mass in vitro, this effect is not cell- and drug-specific because a similar pattern of endogenous Tudor staphylococcal nuclease cleavage is detected in camptothecin-treated HeLa cells
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
NaCl
-
maximum activity of SNR140 and SNR141 SNAse R N-terminal fragments which extends from residues 6 to 140 and 6 to 141 respectively, 0.3 M, studies performed to explore the mechanism of nascent peptide folding
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.00409
-
DNA
-
F34W/W140F mutant, 25C, pH 7.4
0.00431
-
DNA
-
wild type, 25C, pH 7.4
additional information
-
additional information
-
-
-
additional information
-
additional information
-
1250 microg DNA/ml, construct phlbA-sp usp45-nuc, promotor phlbA, lactococcal signal peptide Usp45: sp usp45; 430 microg DNA/ml, construct pusp45-sp usp45-nuc, promoter: pusp45, lactococcal signal peptide Usp45: sp usp45
-
additional information
-
additional information
-
Michaelis-Menten kinetics for wild-type SNase and its mutants, overview
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
-
additional information
-
-
-
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.05
-
3',5'-deoxythymidine diphosphate
-
after 48 h
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
732
-
-
pH 7.4, 25C
1270
-
-
purified recombinant mutant SNase137, pH 7.4, 25C
1319
-
-
purified recombinant mutant SNase139, pH 7.4, 25C
2000
-
-
purified recombinant enzyme, pH and temperature not specified in the publication
2018
-
-
purified recombinant mutant SNase140, pH 7.4, 25C
2298
-
-
purified recombinant mutant SNase141, pH 7.4, 25C
2553
-
-
purified recombinant wild-type SNase, pH 7.4, 25C
additional information
-
-
-
additional information
-
-
antiviral efficacy: in PK-15/Cap-SNase cells and normal PK-15 cells, infected with the classical swine fever virus Shimen strain, the virus titer produced by PK-15/Cap-SNase cells is 100 times lower than that of normal PK-15 cells 5 days after infection, after 6 days, a greater antiviral effect is observed in the PK15/Cap-SNase cells which produced a virus titer that is 3500times lower than the control; the activity of the 5 microl cell lysate containing Cap-SNase is similar to that of 0.5 pg of a standard preparation of SNase, while the linearized plasmid DNA can not be digested when removing Ca2+ by EDTA treatment, indicating that the expressed Cap-SNase retains a good Ca2+-dependent nuclease activity
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.8
-
-
assay at
7
-
-
assay at
7
-
Q0JRI3
assay at
7.4
-
-
assay at
7.4
-
-
assay at
8.8
-
-
1.0 M sodium borate buffer
8.8
-
-
100 mM Ca2+, insolubilized enzyme; 10 mM Ca2+, soluble enzyme
8.8
-
-
assay at
9.5
-
-
10 mM Ca2+, insolubilized enzyme; 1 mM Ca2+, soluble enzyme
10
-
-
1 mM Ca2+, insolubilized enzyme
additional information
-
-
-
additional information
-
-
pH optimum depends on Ca2+ concentration
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
2.5
8
-
the overall secondary and tertiary structure of SNase remains unchanged between pH 2.5 and 8.0
9
10
-
depends on Ca2+ concentration
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Staphylococcus aureus 209P
-
bound
-
Manually annotated by BRENDA team
-
during trophozoite and schizont stages parts of enzyme localized in the cytoplasm
Manually annotated by BRENDA team
Q8VZG7, Q9FLT0
in the interphase and mitosis; in the interphase and mitosis
Manually annotated by BRENDA team
-
the enzyme is secreted
-
Manually annotated by BRENDA team
Staphylococcus aureus V8
-
-
-
-
Manually annotated by BRENDA team
-
mainly nuclear localization
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Staphylococcus aureus (strain MW2)
Staphylococcus aureus (strain MW2)
Staphylococcus aureus (strain MW2)
Staphylococcus aureus (strain MW2)
Staphylococcus aureus (strain MW2)
Staphylococcus aureus (strain MW2)
Staphylococcus aureus (strain MW2)
Staphylococcus aureus (strain MW2)
Staphylococcus aureus (strain MW2)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
12000
-
-
sucrose density gradient centrifugation
17000
-
-
calculation from sequence of amino acid
17000
-
-
molecular weight is calculated by amino acid sequence, the Cap-SNase fusion protein with a molecular weight of 31000 Da is determines by SDS-PAGE and Western blot analysis
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 110000, SDS-PAGE, native protein; x * 95000, SDS-PAGE of the recombinant fusion protein with Maltose Binding Protein
monomer or dimer
-
small globular protein (149 amino acid residues), native protein: no disulfide bond, double mutant (A1C/Q149C): can form disulfide bond
additional information
-
molten globule-like state
additional information
-
structural stability of a P117G variant
additional information
-
structural properties of the enzyme in oligomeric A-forms
additional information
-
four distinct conformational states of wild-type and mutant forms
additional information
-
secondary structure of SNase and its mutants, W140A and Y93W/W140A have less alpha-helical content than the wild-type, overview
additional information
Staphylococcus aureus Foggi
-
four distinct conformational states of wild-type and mutant forms
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
crystal structure of binary Ca2+ and pdTp complexes of the D21E mutant enzyme
-
three-dimensional diffuse x-ray scattering from crystals of the enzyme
-
x-ray structure
-
crystals of the hyperstable mutant enzyme delta+PHS are grown using hanging drop vapor-diffusion methods at 4C from a solution containing 17% 2-methyl-2,4-pentanediol, 2 M CaCl2, 3 M thymine-3',5'-diphosphate, and 25 mM potassium phosphate buffer, pH 8.0
-
crystals of the I92E and I92K variant proteins are obtained at 4C using the hanging-drop vapor-diffusion method
-
E75A mutant: 4C, hanging-drop, precipitating solution 37% (v/v) 2-methyl-2,4-pentanediol and 25 mM potassium phosphate buffer at pH 6.0, protein concentration 9.9 mg/ml before mixing with equal volume of precipitating solution. E75Q: 4C, hanging-drop, precipitating solution 38% (v/v) 2-methyl-2,4-pentanediol and 25 mM potassium phosphate buffer at pH 6.0, starting from 14.2 mg/ml protein. Side chain of His121 is unaffected by elimination of Glu75, histidine moves closer to Glu101 in the structure with E75A. Both crystal structures suggest that the network of polar or ionizable groups connected through hydrogen bonding or charge-charge interactions is a rigid unit, incapable of reorganizing even when strongly stabilizing interactions between Glu75, His124, and Tyr93 are disrupted
-
hanging drop vapor diffusion method
-
LMYKGQPM, short peptide model from staphylococcal nuclease to model the conformational equilibrium between a hairpin conformation and its unfolded state (molecular dynamics simulation), in water, cubic model system, total simulation time 600 ns, starting from a polyproline II conformation, GROMOS96 force field under NVT conditions, 27C: native and non-native hairpins are very close in free energies, interconversion can happen only through the unfolded conformation. Both folding and unfolding events display single exponential kinetics
-
the V66K/P117G/H124L/S128A variant of nuclease is crystallized by the hanging drop vapor diffusion method at 4C, 2 data sets are collected at -173.15C, at pH 7 and 4.7, and the third is collected at 25.15C and pH 5
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3.5
9
-
measurement of stability by two methods: first fluorescence-monitored denaturation with urea, hydrochloric acid and heat. Second by numerical integration of acid titration curves measured potentiometrically under native and unfolding conditions
4.15
-
-
pH-value, at which the acid-denaturation is half completed in wild-type enzyme
4.5
9
-
25C, wild-type enzyme is stable in the range
4.8
-
-
25C, maximal stability of mutant enzyme I92E
9.8
-
-
25C, maximal stability of mutant enzyme I92K
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
15
80
-
temperature range for studying the thermal unfolding transitions
39
-
-
Tm-value of mutant enzyme W140F is 39.1C, Tm-value of mutant enzyme W140Y is 38.6C
42
-
-
Tm-value of mutant enzyme is 41.8C
48
-
-
melting point of mutant enzyme K133A is 47.7C
48
-
-
melting point of the Trp91 insertion mutant enzyme
48.3
-
-
melting point of the Trp15 insertion mutant enzyme
51
-
-
Tm-value for wild-type enzyme is 30.98C
51
-
-
Tm-value of wild-type enzyme is 50.7C
51.7
-
-
melting point of K45C mutant with bound 5,5'-dithiobis-2-nitrobenzoic acid label
52
-
-
melting poit of the wild-type enzyme is 52.32
52.4
-
-
melting point of K45C mutant
53
-
-
melting point of the mutant enzyme DELTA1-141 is 52.8C
53
-
-
melting point of the Trp102 insertion mutant enzyme; melting point of the Trp61 insertion mutant enzyme
53.1
-
-
melting point of wild-type protein
54
-
-
Tm-value without 2-O-alpha-mannosylglycerate is 53.9C
54.5
-
-
melting point of the Trp27 insertion mutant enzyme
56.6
-
-
melting point of the Trp76 insertion mutant enzyme
57
-
-
melting point of the Trp121 insertion mutant enzyme
61
-
-
Tm-value in presence of 0.5 M 2-O-alpha-mannosylglycerate
66
-
-
Tm-value of wild-type enzyme is 66.3C
66.3
-
-
melting point of the wild type enzyme
90
-
-
pH 7.9-8.8, 15 min, 20% loss of activity, pH 3.5 and 9.6, 15 min, 50% loss of activity
additional information
-
-
-
additional information
-
-
heat-stable extracellular enzyme related structurally to a heat-labile cellular nuclease
additional information
-
-
2-O-alpha-mannosylglycerate protects against thermal denaturation. 0.5 M 2-O-alpha-mannosylglycerate increases Tm-value by 7.1C. 0.5 M glycerol or trehalose increase the Tm-value by 0.8C and 4.2C, respectively
additional information
-
-
examination of denaturation (monitored by intrinsic fluorescence of Trp140) due to heat (pH 7): D21N/T33V/T41I/S59A/P117G/A128A mutant(Tm = 72C), K9A mutant (Tm = 71C), E73A mutant (Tm = 67C), E75A mutant (Tm = 70C), E75Q mutant (Tm = 70C), D77A mutant (Tm = 67C), Y91A mutant (Tm = 58C), Y93A mutant (Tm = 52C), E101A mutant (Tm = 71C), K127A mutant (Tm = 75C), A128S mutant (Tm = 71C)
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
2-O-alpha-mannosylglycerate protects against thermal denaturation, 0.5 M 2-O-alpha-mannosylglycerate increases Tm-value by 7.1C. 0.5 M glycerol or trehalose increase the Tm-value by 0.8C and 4.2C, respectively
-
no loss of activity during lyophilization
-
denaturation midpoint for urea is 1.5 M for mutant G20A, 1.1 M urea for mutant G20V, 0.82 Murea or mutant G20I and 2.0 M for wild-type enzyme
-
denaturation of staphylococcal nuclease is induced by guanidinium hydrochloride, wild type and unlabeled mutant proteins are denatured in 20 mM Tris-HCl containing 0.1 M NaCl, pH 7.6 and 10 mM CaCl2, containing various concentrations of guanidinium hydrochloride at 25C for 20 h to reach equilibrium
-
examination of acid-induced denaturation (monitored by intrinsic fluorescence of Trp140) measuring deltaG0H2O: D21N/T33V/T41I/S59A/P117G/A128A mutant: 9.5 kcal/mol, K9A mutant: 6.5 kcal/mol, Y91A mutant: 4.5 kcal/mol, Y91F mutant: 6.7 kcal/mol, Y93A mutant: 3.6 kcal/mol, Y93F mutant: 7.5 kcal/mol, E101A mutant: 8.4 kcal/mol, K127A mutant: 8.9 kcal/mol, A128S mutant: 8.2 kcal/mol. Examination of denaturation (monitored by intrinsic fluorescence of Trp140) with guanidinium chloride measuring the pH at the midpoint of the acid-induced unfolding: D21N/T33V/T41I/S59A/P117G/A128A mutant: 3.05, K9A mutant: 3.16, E73A mutant: 3.21, E75A mutant: 3.28, E75Q mutant: 3.27, D77A mutant: 3.31, Y91A mutant: 3.83, Y93A mutant: 4.10, E101A mutant: 3.14, K127A mutant: 2.94, A128S mutant: 3.20
-
native hairpin conformation is more stable than non-native conformation
-
neural network-based prediction of mutation-induced protein stability changes
-
perchlorate-denatured state has a very high content of secondary structure with no tertiary structure
-
presence of moderate to high concentrations of salt at neutral pH significantly increases thermodinamical stability in both mutant and wild-type enzymes.
-
pressure denaturation of staphylococcal nuclease over a pressure range of 1-3 kilobars at 25C is studied by neutron small-angle scattering and molecular simulation. The globular structure of the enzyme is retained across the folding/unfolding transition although this structure is less compact and elongated relative to the native structure. The findings support a mechanism for the pressure-induced unfolding of the enzyme in which water penetration into the hydrophobic core plays a central role
-
OXIDATION STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ongoing denaturation by exposure to oxidative stress brought on by illumination of a solution containing 0.145 mM enzyme and 0.5 mM fullerol with polychromatic white light
-
681601
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant protein is purified by anion exchange chromatography on a Q Sepharose Fast Flow column using a linear gradient of NaCl (0-500 mM) in 20 mM Tris-HCl at pH 8.0, 2 mM dithiothreitol and 10% glycerol
Q8VZG7, Q9FLT0
recombinant protein is purified by anion exchange chromatography on a Q Sepharose Fast Flow column using a linear gradient of NaCl (0-500 mM) in 20 mM Tris-HCl at pH 8.0, 2 mM dithiothreitol and 10% glycerol
-
recombinant protein is purified by anion exchange chromatography on a Q Sepharose Fast Flow column using a linear gradient of NaCl (0-500 mM) in 20 mM Tris-HCl at pH 8.0, 2 mM dithiothreitol and 10% glycerol
Q0JRI3
recombinant fusion protein with Maltose Binding Protein
-
proline free mutant
-
recombinant enzyme
-
recombinant enzyme from Lactococcus lactis cell culture medium by cation exchange chromatography
-
recombinant protein
-
recombinant proteins
-
10 mM DTT added to expose sulfhydryl groups, and the solution passes through a short column of PD-10 to remove the reducing agent
-
recombinant wild-type and mutant enzymes from Escherichia coli
-
wild type and mutant enzyme are purified by a series of urea extraction, ethanol precipitation, and ion exchange chromatography
-
wild type and mutant enzymes are purified by a chelating-sepharose fast flow column
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Escherichia coli
Q8VZG7, Q9FLT0
expression in Escherichia coli
-
cloning of the full-length cDNA from Picea abies encoding Tudor staphylococcal nuclease, expression in Escherichia coli
Q0JRI3
expression of protein fragment results in the formation of insoluble inclusion bodies, successful expression as fusion protein with Maltose Binding Protein
-
possible target for the anti-malarial therapy tested by RNAi. Role of PfTSN at asexual blood stages investigated by treatment of synchronized parasites at late ring stage with siRNA (analysed by manual counting and by morphological examination 40 h after treatment, additionally analyzed by real time PCR and immunofluoresence), 50 microg/ml concentration of siRNA: over 50% reduction in parasitemia observed, 100 microg/ml concentration of PfTSN siRNA: 60-70% reduction in parasitemia. Effect of PfTSN siRNA over the course of erythrocytic cycle investigated by treatment of parasites with PfTSN siRNA at late ring stage and morphologically examined after the treatment: after 2-3 hours abnormalities (vacuolation, size of vacuols seems to increase over time), after 12 hours most parasites are dead
-
a plasmid pcDNA-Cap-SNase is constructed for expressing a fusion protein of classical swine fever virus capsid Cap and staphylococcal nuclease, a mammalian cell line PK-15 expressing stably the fusion protein Cap-SNase
-
expressed in Escherichia coli
-
expression of poly-his-nuclease R in Escherichia coli
-
expression of wild type and mutant proteins in Escherichia coli BL21(DE3), formation of inclusion bodies
-
gene nuc1, recombinant expression in Escherichia coli strain BL21(DE3)
-
generation of Vibrio anguillarum ghost by coexpression of PhiX 174 Lysis E gene and SNA gene. Gene fragment encoding SNA amplified by PCR using Staphlococcus aureus genomic DNA. construction of plasmid pRK-lambda-P(R)-cI-SNA. Construction of dual vector expressing PhiX 174 lysis E gene and SNA: pRK-kP(R)-cI-E-SNA. Transformation of Escherichia coli SM10-lambda-pir used as donor for plasmid transfer with Vibrio anguillarum via conjugation. Induction of protein expression by temperature elevation
-
proline free mutant
-
purification of DNA from the cell-associated herpesvirus Marek's disease virus. 150 U of Micrococcal nuclease added to gallid herpesvirus type 2 virus infected cells (GaHV-2 strain 648A) in 100 microl reaction volume, digestion followed by PEG precipitation yields high-molecular weight DNA of greater than 75% pure GaHV-2 DNA suitability for both direct pyrosequencing and further amplification using isothermal polymerase
-
recombinant Trp insertion enzymes
-
staphylococcal nuclease fused at its N-terminus to signal peptide of the lactococcal Usp45 protein (SP Usp45-NucB), as reporter for expression and secretion in Lactobacillus bulgaricus, SDS PAGE and western blot of culture supernatant and cell lysate used for analysis
-
the enzyme is cloned into an expression-secretion vector lacking its own signal peptide but being fused to a lactococcal sequence encoding a signal peptide. It is then fused to a lactococcal sequence encoding a signal peptide. Functional recombinant expression of the enzyme under the control of a lactococcal promoter, that is inducible by zinc starvation, in Lactococcus lactis subsp. cremoris model strain MG1363, the enzyme is secreted to the GM17v culture medium, expression method optimization, overview
-
a hyperstable, acid-resistant form of SNase known as delta+PHS is expressed in Escherichia coli BL21/DE3 cells
-
BHL-21 cells stably express staphylococcal nuclease fused to dengue 2 virus capsid protein for CTVI. The intracellular expressed fusion protein is correctly folded and has no cytotoxicity on the mammalian host cells
-
construction and co-expression of Staphylococcal nuclease in Escherichia coli
-
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)/pLysS
-
gene insert cloned into pET24a+plasmid, transformation of BL21(DE3) Escherichia coli cells for expression
-
overexpression of wild-type and mutant enzymes in Escherichia coli
-
recombinant wild type protein and mutants are expressed in Escherichia coli
-
wild type and mutant Snases are expressed in Escherichia coli BL-21 (DE3) as inclusion bodies
-
wild type enzyme and the mutants K28C/H124C K28C/K97C are expressed in BL21 (DE3) and K28C/K97C in BL21 (DE3) Escherichia coli cells
-
wild-type and S28G mutant expressed in Escherichia coli strain AR120
-
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
down-regulation of enzyme using interfering RNA results in pollen degeneration and a dramatic reduction in plant fertility, providing further evidence for an essential role of TSN in plant reproduction
Q8VZG7, Q9FLT0
in hyperplasia specimens and normal epithelium, protein is weakly or negatively expressed
-
knocked down staphylococcal nuclease domain-containing protein 1 in vitro with small interfering RNA causes a significant decrease in cell growth
-
Tudor staphylococcal nuclease knockdown leads to activation of ectopic cell death during reproduction, impairing plant fertility, HeLa cells transfected with Tudor staphylococcal nuclease short interfering RNA show a dramatic increase in apoptotic response to camptothecin accompanied by a 7.9fold increase in activation of caspase-3 and increased cleavage of PARP and lamin-A (by 3.7fold and 6.7fold, respectively), moreover, reduction of Tudor staphylococcal nuclease levels induces apoptosis even in the absence camptothecin, leading to 7.7fold, 6.1fold and 11fold increases in apoptotic markers, demonstrating that Tudor staphylococcal nuclease is indispensable for the maintenance of cell viability
-
staphylococcal nuclease domain-containing protein 1 is highly expressed in recurrent androgen-insensitive prostate cancer tissues, protein expression intensity increases with increasing grade and aggressiveness of the cancer, staphylococcal nuclease domain-containing protein 1 mRNA is highly expressed in the cytoplasm of cancer cells but is negative to weak in noncancerous cells
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
D790E
-
by site-directed mutagenesis, proteolysis of Tudor staphylococcal nuclease does not occur either when the P1 position of the DAVD motif is mutated or after treatment with the pan-caspase inhibitor zVAD-fluoromethylketone, expression of mutant under normal conditions enhances cell proliferation in both cancer (HeLa) and non-cancer (HEK-293) cells compared with mock- and wild-type Tudor staphylococcal nuclease-transfected samples, under camptothecin-induced apoptosis, expression of Tudor staphylococcal nuclease mutant results in a 35% increment in viable HeLa cells, suggesting that caspase-mediated proteolysis of enzyme is important for the progression of apoptosis
DELTA140-149
-
deletion of the 10 C-terminal residues, mutant proteins are in a non-native or disordered state under physiological conditions, folding is induced by addition of an inhibitor or substrate
G79S/H124LC80-C116
-
effects on the stability and conformation of the folded protein
H124LC77-C118
-
effects on the stability and conformation of the folded protein
H124LC79-C118
-
effects on the stability and conformation of the folded protein
H124LC80-C116
-
effects on the stability and conformation of the folded protein
INS33A34
-
insertion of an alanine between residues 33 and 34, mutant proteins are in a non-native or disordered state under physiological conditions, folding is induced by addition of an inhibitor or substrate
K45C
-
insertion of a cysteine to enable labeling with thiol reactive ligands, e.g. 5,5'-dithiobis-2-nitrobenzoic acid, CD-spectra of wild type enzyme, mutant and mutant with 5,5'-dithiobis-2-nitrobenzoic acid label indicate, that the protein have very similar secondary structures
P11A/P31A/P42A/P47T/P56A/P117G
-
proline free mutant, conformationally different from wild type protein, 1.4% of wild type activity
P47G/P117G/H124L/W140H
-
tryptophan-free mutant used for the insertion of a unique tryptophan at positions 15, 27, 61, 76, 91, 102, and 121, mutant enzymes used to study the enzyme folding kinetics, variants are destabilized but maintain the ability to refold in the native-like structure
T62C
-
designed for the insertion of a cysteine reactive label
T62P
P00644
highly destabilized variant of enzyme, exists in the unfolded state over a wide pH-range, can be fully refolded to the native folding by addition of osmolytes
A128S
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to study inductive effects and longer-range interactions between elements of the network, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy)
A1C/Q149C
-
SNase double mutant, N- and C-terminal residues replaced by cysteine, constructed from the plasmid (pMT7-SN) of wild-type SNase using the Kunkel method, can form disulfide bond
A90S
-
pH-value, at which the acid-denaturation is half completed is 4.19, compared to pH 3.76 for wild-type enzyme. The apparent number of protons which trigger the denaturation and are taken up by the protein upon denaturation is 1.0 for the mutant enzyme compared to 1.8 for wild-type enzyme
D143G
-
Tm value for mutant enzyme is 50.53C, compared to 50.98C for wild-type enzyme
D143K
-
charge reversal
D143N
-
charge reversal
D146G
-
Tm value for mutant enzyme is 50.99C, compared to 50.98C for wild-type enzyme
D19G
-
Tm value for mutant enzyme is 52.06C, compared to 50.98C for wild-type enzyme
D19K
-
charge reversal
D19L
-
charge reversal
D21G
-
Tm value for mutant enzyme is 53.74C, compared to 50.98C for wild-type enzyme
D21K
-
charge reversal
D21N
-
charge reversal
D21N/T33V/T41I/S59A/P117G/A128A
-
hyperstable engineered form of staphylococcal nuclease (SNase)
D40G
-
Tm value for mutant enzyme is 50.44C, compared to 50.98C for wild-type enzyme
D77A
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to study inductive effects and longer-range interactions between elements of the network, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy) , determination of tautomeric states of His121 and His124, pH near pI
D77G
-
loss of catalytic efficiency of 16% and thermal stabilty of 26% relative to the wild-type protein
D77G
-
Tm value for mutant enzyme is 44.14C, compared to 50.98C for wild-type enzyme
D77K
-
charge reversal
D77N
-
charge reversal
D77N
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to study inductive effects and longer-range interactions between elements of the network, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy) , determination of tautomeric states of His121 and His124, pH near pI
D83G
-
Tm value for mutant enzyme is 37.21C, compared to 50.98C for wild-type enzyme
D95G
-
Tm value for mutant enzyme is 37.38C, compared to 50.98C for wild-type enzyme
DELTA1-139
-
mutant lacks tertiary structure, fluorescence of the mutant is much lower than that of the wild-type enzyme
DELTA1-141
-
intact tertiary conformation, melting point is nearly identical to wild-type enzyme
E101A
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to examine shortrange effects on His124, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy)
E101G
-
Tm value for mutant enzyme is 43.04C, compared to 50.98C for wild-type enzyme
E10G
-
Tm value for mutant enzyme is 43.8C, compared to 50.98C for wild-type enzyme
E10K
-
charge reversal
E10Q
-
charge reversal
E122G
-
Tm value for mutant enzyme is 44.12C, compared to 50.98C for wild-type enzyme
E122K
-
charge reversal
E122Q
-
charge reversal
E129G
-
Tm value for mutant enzyme is 34.59C, compared to 50.98C for wild-type enzyme
E135G
-
Tm value for mutant enzyme is 44.54C, compared to 50.98C for wild-type enzyme
E135K
-
charge reversal
E135Q
-
charge neutralization
E142G
-
Tm value for mutant enzyme is 49.41C, compared to 50.98C for wild-type enzyme
E43G
-
Tm value for mutant enzyme is 54.99C, compared to 50.98C for wild-type enzyme
E52G
-
Tm value for mutant enzyme is 52.1C, compared to 50.98C for wild-type enzyme
E57G
-
Tm value for mutant enzyme is 46.6C, compared to 50.98C for wild-type enzyme
E57K
-
charge reversal
E57Q
-
charge reversal
E67G
-
Tm value for mutant enzyme is 46.53C, compared to 50.98C for wild-type enzyme
E67Q
-
charge reversal
E73A
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to study inductive effects and longer-range interactions between elements of the network, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy)
E73G
-
loss of catalytic efficiency of 24% and thermal stabilty of 22% relative to the wild-type protein
E73G
-
Tm value for mutant enzyme is 37C, compared to 50.98C for wild-type enzyme
E73G/D77G
-
loss of thermal stabilty of 47% relative to the wild-type protein
E73G/E75G
-
loss of thermal stabilty of 59% relative to the wild-type protein
E73G/E75G/D77G
-
loss of thermal stabilty of 65% relative to the wild-type protein
E73K
-
charge reversal
E73Q
-
charge reversal
E75A
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to examine short-range (up to 6.4 Angstrom) Coulomb and hydrogen bonding effects on His121, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy), determination of tautomeric states of His121 and His124, pH near pI
E75G
-
Tm value for mutant enzyme is 36.99C, compared to 50.98C for wild-type enzyme
E75G/D77G
-
loss of thermal stabilty of 58% relative to the wild-type protein
E75K
-
charge reversal
E75Q
-
charge reversal
E75Q
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to examine short-range (up to 6.4 Angstrom) Coulomb and hydrogen bonding effects on His121, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy)
F34W/W140F
-
characterized by far and near UV CD, gel filtration, ANS-binding fluorescence, enzymatic parameters are similar to those of the wild type, similar substrate affinity to the wild type enzyme
F61W/W140A
-
the mutant shows reduced activity with higher Michaelis-Menten constants, Km, and lower maximum reaction rate compared to the wild-type enzyme, the mutant also shows a more rapid loss of secondary and tertiary structure by Gdn-HCl unfolding than the wild-type enzyme
F76W/W140H
-
mutation causes decrease in thermal stability
G20A
-
2% of wild-type activity. Km(Ca) is almost 20fold higher than the wild-type enzyme. Denaturation midpoint for the mutant enzyme is 1.5 M urea compared to 2.0 M urea for the wild-type enzyme
G20I
-
0.21% of wild-type activity. Km(Ca) is almost 50fold higher than the wild-type enzyme. Denaturation midpoint for mutant enzyme is 0.82 M urea compared to 2.0 M urea for the wild-type enzyme
G20V
-
0.45% of wild-type activity. Km(Ca) is almost 20fold higher than the wild-type enzyme. Denaturation midpoint for the mutynt enzyme is 1.1 M urea compared to 2.0 M urea for the wild-type enzyme
G50F/V51N/P117G/H124L/S128A/DELTA44-49
-
hyperstable, acid-resistant mutant form of enzyme known as delta+PHS
G79S
-
pH-value, at which the acid-denaturation is half completed is 4.39, compared to pH 3.76 for wild-type enzyme. The apparent number of protons which trigger the denaturation and are taken up by the protein upon denaturation is 1.4 for the mutant enzyme compared to 1.8 for wild-type enzyme
G88V
-
pH-value, at which the acid-denaturation is half completed is 3.57, compared to pH 3.76 for wild-type enzyme. The apparent number of protons which trigger the denaturation and are taken up by the protein upon denaturation is 3.0 for the mutant enzyme compared to 1.8 for wild-type enzyme
H124E
-
charge reversal
H124L
-
pH-value, at which the acid-denaturation is half completed is 2.98, compared to pH 3.76 for wild-type enzyme. The apparent number of protons which trigger the denaturation and are taken up by the protein upon denaturation is 2.8 for the mutant enzyme compared to 1.8 for wild-type enzyme
H124Q
-
charge neutralization
I92E
-
wild-type enzyme exhibits a broad range of pH-independence from pH 4.5 to pH 9, mutant enzyme exhibits pronounced pH-dependence with a maximal stability at pH 4.9
I92K
-
wild-type enzyme exhibits a broad range of pH-independence from pH 4.5 to pH 9.0, mutant enzyme exhibits pronounced pH-dependence with a maximal stability at pH 9.8
K127A
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to examine shortrange effects on His124, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy)
K127E
-
charge reversal
K127Q
-
charge neutralization
K133A
-
intact tertiary conformation, melting point is 4.6C lower than that of the wild-type enzyme
K133Q
-
charge neutralization
K28C/H124C
-
generated by site-directed mutagenesis, native and non-native conformations are observed, and the non-native conformation expands with increasing guanidinium hydrochloride concentrations, the non-native chains of the derivative exhibits different changes of persistence length at higher guanidinium hydrochloride concentrations, suggesting a subdomain-specific collapse of the denatured state of SNase, this local chain specific collapse is likely to play a role in modulating the formation of early intermediate during protein folding
K28C/K97C
-
generated by site-directed mutagenesis, native and non-native conformations are observed, and the non-native conformation expands with increasing guanidinium hydrochloride concentrations, the non-native chains of the derivative exhibits different changes of persistence length at higher guanidinium hydrochloride concentrations, suggesting a subdomain-specific collapse of the denatured state of SNase, this local chain specific collapse is likely to play a role in modulating the formation of early intermediate during protein folding
K28E
-
charge reversal
K28Q
-
charge neutralization
K48E
-
charge reversal
K48Q
-
charge neutralization
K63E
-
charge reversal
K63Q
-
charge neutralization
K64E
-
charge reversal
K64Q
-
charge neutralization
K70E
-
charge reversal
K70Q
-
charge neutralization
K78E
-
charge reversal
K78Q
-
charge neutralization
K84E
-
charge reversal
K84Q
-
charge neutralization
K97E
-
charge reversal
K97Q
-
charge neutralization
K9A
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to study inductive effects and longer-range interactions between elements of the network, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy)
S28C
-
this mutant contains a five-amino acid type I beta-turn from concanavalin A in place of residues 27-30 of SNase
V66D/P117G/H124L/S128A
-
production by site-directed mutagenesis, pka value shifts to 7.79 and, after chemical denaturation, to 8.05
V66D/P117G/H124L/S128A/G50F/V51N/DELTA44-49
-
production by site-directed mutagenesis, pka value shifts to 8.95 and, after chemical denaturation, to 8.73
V66E/P117G/H124L/S128A
-
production by site-directed mutagenesis, pka value shifts to 8.80 and, after chemical denaturation, to 8.99
V66E/P117G/H124L/S128A/G50F/V51N/DELTA44-49
-
production by site-directed mutagenesis, pka value shifts to 9.07 and, after chemical denaturation, to 8.80
V66K
-
production by site-directed mutagenesis, pka value shifts to 6.38 after chemical denaturation
V66K
-
the substitution affects the H/D exchange properties of the protein globally, even when Lys66 is neutral. The conformational dynamics of the protein probed by H/D exchange indicate that, while both the global fluctuation and the local fluctuation are increased by the substitution, the global fluctuations are enhanced by protonation of Lys-66
V66K/P117G/H124L/S128A
-
production by site-directed mutagenesis, pka value shifts to 6.35
V66K/P117G/H124L/S128A/G50F/V51N/DELTA44-49
-
production by site-directed mutagenesis, pka value shifts to 5.63 and, after chemical denaturation, to 5.83
V66L
-
pH-value, at which the acid-denaturation is half completed is 3.36, compared to pH 3.76 for wild-type enzyme. The apparent number of protons which trigger the denaturation and are taken up by the protein upon denaturation is 2.6 for the mutant enzyme compared to 1.8 for wild-type enzyme
V66L/G79S/G88V
-
pH-value, at which the acid-denaturation is half completed is 3.67, compared to pH 3.76 for wild-type enzyme. The apparent number of protons which trigger the denaturation and are taken up by the protein upon denaturation is 1.1 for the mutant enzyme compared to 1.8 for wild-type enzyme
V66L/G88V
-
pH-value, at which the acid-denaturation is half completed is 3.42, compared to pH 3.76 for wild-type enzyme. The apparent number of protons which trigger the denaturation and are taken up by the protein upon denaturation is 1.6 for the mutant enzyme compared to 1.8 for wild-type enzyme
W140A
-
mutant lacks tertiary structure, fluorescence of the mutant is much lower than that of the wild-type enzyme
W140A
-
DNA hydrolysis activity is 70% of wild-type activity
W140A
-
the mutant shows reduced activity with higher Michaelis-Menten constants, Km, and lower maximum reaction rate compared to the wild-type enzyme, the mutant also shows a more rapid loss of secondary and tertiary structure by Gdn-HCl unfolding than the wild-type enzyme
W140C
-
DNA hydrolysis activity is 75% of wild-type activity
W140D
-
DNA hydrolysis activity is 65% of wild-type activity
W140E
-
DNA hydrolysis activity is 65% of wild-type activity
W140F
-
mutation causes decrease in thermal stability
W140F
-
native-like structure and native-like activity under physiological conditions
W140G
-
DNA hydrolysis activity is 75% of wild-type activity
W140H
-
mutation causes decrease in thermal stability
W140H
-
native-like structure and native-like activity under physiological conditions
W140I
-
DNA hydrolysis activity is 70% of wild-type activity
W140K
-
DNA hydrolysis activity is 70% of wild-type activity
W140L
-
mutation causes decrease in thermal stability
W140L
-
DNA hydrolysis activity is 75% of wild-type activity
W140M
-
DNA hydrolysis activity is 70% of wild-type activity
W140N
-
DNA hydrolysis activity is 75% of wild-type activity
W140P
-
DNA hydrolysis activity is 55% of wild-type activity
W140Q
-
DNA hydrolysis activity is 75% of wild-type activity
W140R
-
DNA hydrolysis activity is 75% of wild-type activity
W140S
-
DNA hydrolysis activity is 75% of wild-type activity
W140T
-
DNA hydrolysis activity is 75% of wild-type activity
W140V
-
DNA hydrolysis activity is 75% of wild-type activity
W140Y
-
mutation causes decrease in thermal stability
W140Y
-
native-like structure and native-like activity under physiological conditions
Y54C/I139C
-
production by site-directed mutagenesis, the oxidized form assumes a more compact denatured structure under acidic conditions than the wild type, the kinetic measurements reveal that the refolding reactions of both the reduced and oxidized forms of mutant are similar to those of the wild type protein
Y54C/I139C/DELTA140-149
-
production by site-directed mutagenesis, under physiological conditions, the reduced form appears to assume a denatured structure, in contrast, the oxidized form forms a native-like structure
Y91A
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to examine short-range (up to 6.4 Angstrom) Coulomb and hydrogen bonding effects on His121, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy)
Y91F
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to examine short-range (up to 6.4 Angstrom) Coulomb and hydrogen bonding effects on His121, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy) , determination of tautomeric states of His121 and His124, pH near pI
Y93A
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to examine short-range (up to 6.4 Angstrom) Coulomb and hydrogen bonding effects on His121, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy) , determination of tautomeric states of His121 and His124, pH near pI
Y93F
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to examine short-range (up to 6.4 Angstrom) Coulomb and hydrogen bonding effects on His121, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy) , determination of tautomeric states of His121 and His124, pH near pI
Y93W/W140A
-
the mutant shows reduced activity with higher Michaelis-Menten constants, Km, and lower maximum reaction rate compared to the wild-type enzyme, the mutant also shows a more rapid loss of secondary and tertiary structure by Gdn-HCl unfolding than the wild-type enzyme
L25A
-
pH-value, at which the acid-denaturation is half completed is 4.15, compared to pH 3.76 for wild-type enzyme. The apparent number of protons which trigger the denaturation and are taken up by the protein upon denaturation is 1.2 for the mutant enzyme compared to 1.8 for wild-type enzyme
additional information
-
almost all mutations are destabilizing, the average loss of stability for all of the charge neutralization substitutions is 0.5 kcal/mol and the average loss of stability for all of the charge reversal substitutions is 1.0 kcal/mol
additional information
-
generation by site-directed mutagenesis of C-terminal truncated SNases, i.e. SNase137, SNase139, SNase140, and SNase141 containing residues 1-137, 1-139, 1-140, and 1-141, respectively. The mutants show reduced activities compared to the wild-type enzyme. Determination of the secondary structures of the four SNase fragments, overview
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
effect of denaturants at low concentrations on very unstable mutant enzyme forms
-
reversible thermal denaturation
-
high concentration of urea and guanidinium chloride denaturation is completely reversible at pH 3.5-3.7, 55-65C
-
refolding of acid-unfolded enzyme induced anions
-
atomic force microscopy, unfolding experiment: multiplicity of unfolding intermediates of SNase at the single-molecule level, mechanical unfolding pathways can be changed drastically under the acid-denatured condition (native conditions: 1 mM EGTA, pH 8.0, acid-denaturated condition: 1 mM EGTA, pH 2.5) or in the presence of ligands (Ca2+) and inhibitors (2'-deoxythymidine 3',5'-diphosphate)
-
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
medicine
-
as staphylococcal nuclease domain-containing protein 1 mRNA is overexpressed in cancer cells, the growth of these cells is suppressed following staphylococcal nuclease domain-containing protein 1 knock-down in vitro, thus representing a promising prostate cancer biomarker and therapeutic target, evidence for the diagnostic potential of staphylococcal nuclease domain-containing protein 1 in prostate surgical specimens equivalent or better than that of alpha-methylacyl-CoA racemase
drug development
-
attractive potential target for the development of anti-malarial drugs
medicine
-
possible target for the anti-malarial therapy tested by RNAi
analysis
-
staphylococcal nuclease fused at its N-terminus to signal peptide of the lactococcal Usp45 protein (SP Usp45-NucB), as reporter for expression and secretion in Lactobacillus bulgaricus
analysis
-
purification of DNA from the cell-associated herpesvirus Mareks disease virus
analysis
-
the enzyme is a useful tool for mapping chromatin structure in eukaryotes, usage of the enzyme to determine the positions of nucleosomes within a region of DNA to identify dynamic changes induced during gene regulation
drug development
-
generation of Vibrio anguillarum ghost by coexpression of PhiX 174 Lysis E gene and SNA gene, potential vaccine for fishes against vibriosis
medicine
-
capsid-targeted viral inactivation as an antiviral strategy against classical swine fever infection, the fusion protein Cap-SNase can inhibit effectively the production of classical swine fever virus, resulting in a reduction in infectious titers
molecular biology
-
the enzyme is a useful tool for mapping chromatin structure in eukaryotes, usage of the enzyme to determine the positions of nucleosomes within a region of DNA to identify dynamic changes induced during gene regulation
molecular biology
-
purified enzyme can be used as an exogenous reagent to clear cellular extracts and improve protein purification
drug development
Staphylococcus aureus KCCM 11335
-
generation of Vibrio anguillarum ghost by coexpression of PhiX 174 Lysis E gene and SNA gene, potential vaccine for fishes against vibriosis
-
analysis
-
peptide model LMYKGQPM from staphylococcal nuclease can serve as model for faster folding beta-hairpins pursuing fast folding events
biotechnology
-
co-expression of Staphylococcal nuclease in Escherichia coli to reduce the viscosity of the bioprocess feedstock through auto-hydrolysis of nucleic acids, viscosity is an important physical property of the process stream and a significant factor in the optimization of various downstream processing unit operations including cell disruption, clarification, filtration, and chromatography