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 hide
3.1.31.1
-
RECOMMENDED NAME
GeneOntology No.
micrococcal nuclease
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
endonucleolytic cleavage to nucleoside 3'-phosphates and 3'-phosphooligonucleotide end-products
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
CAS REGISTRY NUMBER
COMMENTARY hide
9013-53-0
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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
-
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
209P
-
-
Manually annotated by BRENDA team
Staphylococcus aureus CCTCC AB91093
-
-
-
Manually annotated by BRENDA team
strain KCCM 11335, Korean Culture Center of Microorganisms, Korea
-
-
Manually annotated by BRENDA team
gene nuc1
-
-
Manually annotated by BRENDA team
a community-acquired CA-MRSA strain, gene nuc
-
-
Manually annotated by BRENDA team
strain V8
-
-
Manually annotated by BRENDA team
gene nucM
A0A076JSR1
UniProt
Manually annotated by BRENDA team
gene nucM
UniProt
Manually annotated by BRENDA team
gene nucM
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
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
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
-
-
?
methyl-pdTp-nitrophenyl + H2O
?
show the reaction diagram
-
-
-
-
?
nitrophenyl-pdT + H2O
?
show the reaction diagram
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
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
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
DNA + H2O
3'-deoxymononucleotides + dinucleotides
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
Q7A5U0, Q7A6P2
activates slightly at 5 mM
Cu2+
-
minimal activation if Ca2+ is replaced by Cu2+
Fe2+
-
minimal activation if Ca2+ is replaced by Fe2+
Ni2+
Q7A5U0, Q7A6P2
activates slightly at 0.05 mM
Sr2+
-
DNase but no RNase activity if Ca2+ is replaced Sr2+
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
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
Ca2+
-
competitive
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%
-
Co2+
Q7A5U0, Q7A6P2
isozyme Nuc1 shows 79% activity at 5 mM concentration
Cu2+
-
competitive
deoxythymidine 3',5'-bisphosphate
deoxythymidine 3',5'-diphosphate
-
in the presence of pdTp, unfolding forces increase drastically
Hg2+
-
competitive
metacaspase mcII-Pa
Mn2+
Q7A5U0, Q7A6P2
isozyme Nuc1 activity drops sharply at 5 mM concentration; isozyme Nuc2 activity decreases with an increase in Mn2+ concentration
Mononucleotides
-
with 5'-phosphate end group
oligonucleotides
-
with 5'-phosphate end group
thymidine 3',5'-bisphosphate
-
competitive
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mecaptoethanol
Q7A5U0, Q7A6P2
;
-
dithiothreitol
Q7A5U0, Q7A6P2
-
DTT
Q7A5U0, Q7A6P2
163.4% of initial isozyme Nuc1 activity at 1 mM
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
Triton X-100
Q7A5U0, Q7A6P2
; 210.5% of initial isozyme Nuc1 activity at 1%
Tween-20
Q7A5U0, Q7A6P2
;
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00409 - 0.00431
DNA
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
Staphylococcus aureus
-
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.05
3',5'-deoxythymidine diphosphate
Plasmodium falciparum
-
after 48 h
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
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
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.9
-
assay at
6.8
-
assay at
7.5
-
assay at
9.5
-
10 mM Ca2+, insolubilized enzyme; 1 mM Ca2+, soluble enzyme
additional information
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2.5 - 8
-
the overall secondary and tertiary structure of SNase remains unchanged between pH 2.5 and 8.0
3 - 10
Q7A5U0, Q7A6P2
effects of pH on thermonuclease activity of isozymes Nuc1 and Nuc2, profile overview; effects of pH on thermonuclease activity of isozymes Nuc1 and Nuc2, profile overview
9 - 10
-
depends on Ca2+ concentration
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
assay at
30
Q7A5U0, Q7A6P2
assay at; assay at
50
Q7A5U0, Q7A6P2
recombinant enzyme
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 80
Q7A5U0, Q7A6P2
effects of temperature on thermonuclease activity of isozymes Nuc1 and Nuc2, profile overview; effects of temperature on thermonuclease activity of isozymes Nuc1 and Nuc2, profile overview
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
high SND1 enzyme expression level
Manually annotated by BRENDA team
additional information
-
expression of the enzyme and of angiotensin II type 1 receptor are positively correlated
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
mainly nuclear localization
Manually annotated by BRENDA team
additional information
-
human Tudor staphylococcal nuclease interacts with the Ras-GAP SH3 domain-binding protein and is recruited into stress granules, the main type of discrete RNA-containing cytoplasmic foci structure that is formed under stress conditions
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Staphylococcus aureus (strain COL)
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)
Staphylococcus aureus (strain MW2)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
12000
-
sucrose density gradient centrifugation
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
-
1 * 16800
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
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure analysis, PDB ID 2SNS
-
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
ORGANISM
UNIPROT
COMMENTARY hide
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
657910
4.15
-
pH-value, at which the acid-denaturation is half completed in wild-type enzyme
664352
4.5 - 9
-
25C, wild-type enzyme is stable in the range
666025
4.8
-
25C, maximal stability of mutant enzyme I92E
666025
5 - 10
-
recombinant mutant enzymes, stable, overview
729399
9.8
-
25C, maximal stability of mutant enzyme I92K
666025
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
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
48.3
-
melting point of the Trp15 insertion mutant enzyme
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
53.1
-
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
75
Q7A5U0, Q7A6P2
1 h, heat-treated isozyme Nuc1 is still able to degrade the DNA substrate; 1 h, heat-treated isozyme Nuc2 is still able to degrade the DNA substrate
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
GENERAL STABILITY
ORGANISM
UNIPROT
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
-
correlation between the magnitude of protein stabilization and the restriction of fast backbone motions, mannosylglycerate restricts local motions in addition to the global motions of the protein. Unfolding/folding pathway remain undisturbed in the presence of mannosylglycerate but the solute shows a specific effect on the local motions of beta-sheet residues
-
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
-
enzyme tryptophan fluorescence spectra, fluorescence measurements of protein unfolding under pressure, high-pressure fluorescence spectroscopy and single value decomposition analysis, overview
-
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
-
no loss of activity during lyophilization
-
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
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
LITERATURE
10 mM DTT added to expose sulfhydryl groups, and the solution passes through a short column of PD-10 to remove the reducing agent
-
proline free mutant
-
recombinant enzyme
-
recombinant enzyme from Lactococcus lactis cell culture medium by cation exchange chromatography
-
recombinant fusion protein with Maltose Binding Protein
-
recombinant GST-tagged enzyme constructs from Escherichia coli strain Rossetta (DE3) by glutathione affinity chromatography, and anion or cation exchange chromatography, followed by dialysis
recombinant His-tagged truncated isozyme Nuc2 lacking the N-terminal membrane anchor by nickel-affinity chromatography
Q7A5U0, Q7A6P2
recombinant protein
-
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 proteins
-
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
LITERATURE
a hyperstable, acid-resistant form of SNase known as delta+PHS is expressed in Escherichia coli BL21/DE3 cells
-
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
-
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
-
cloning of the full-length cDNA from Picea abies encoding Tudor staphylococcal nuclease, expression in Escherichia coli
construction and co-expression of Staphylococcal nuclease in Escherichia coli
-
expressed in Escherichia coli
-
expression and interaction analysis of the enzyme fused to a GAL4-DNA-binding domain and the three Argonaute proteins in the Saccharomyces cerevisiae strain Y187 two-hybrid system, expression of N-terminally GST-tagged enzyme constructs comprising amino acid residues 1-889, 1-660, and residues 320-889, in Escherichia coli strain Rossetta (DE3)
expression in Escherichia coli
expression in Hep-3B cells
-
expression of poly-his-nuclease R in Escherichia coli
-
expression of protein fragment results in the formation of insoluble inclusion bodies, successful expression as fusion protein with Maltose Binding Protein
-
expression of the enzyme from a plsmid encoding RFP-epitope-tagged Tudor-SN in HeLa cells, localization profile of the Tudor-SN-AGTR1-3'UTR complex, GFP-MS2 fluorescence and immuno-labeling analysis, overview
-
expression of wild type and mutant proteins in Escherichia coli BL21(DE3), formation of inclusion bodies
-
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
-
gene nuc1, recombinant expression in Escherichia coli strain BL21(DE3)
-
gene nuc1, sequence comparison and phylogenetic analysis, functional recombinant expression in Escherichia coli strain BL21(DE3); gene nuc2, sequence comparison and phylogenetic analysis, functional recombinant expression in Escherichia coli strain BL21(DE3)
Q7A5U0, Q7A6P2
gene nuc1, sequence comparison, recombinant expression of C-terminally His6-tagged isozyme Nuc1 in Escherichia coli strain strain ER2566; gene nuc2, sequence comparison, the portion of nuc2 encoding amino acids N26-K177 is amplified from AH1263 genomic DNA, recombinant expression of the truncated version as C-terminally His6-tagged isozyme Nuc2, and expression of isozyme Nuc2 as fusion proteins Nuc2-GFP and Nuc2-PhoA, respectively, in Escherichia coli strain strain ER2566, creating strain AH2591. Expression of recombinant chimeric His-tagged fusion proteins in Staphylococcus aureus nuc-deficient mutant strain AH1680
Q7A5U0, Q7A6P2
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
-
overexpression of wild-type and mutant enzymes in Escherichia coli
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
-
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
-
recombinant wild type protein and mutants are expressed in Escherichia coli
-
SaeRS is required for expression of the nuc gene, expression from plasmid containing the nuc promoter coupled to sGFP (pCM20), transformed into the Staphylococcus aureus USA300 wild-type-LAC and sae, agr, and sigB mutant strains
-
stable enzyme overexpression in hepatocellular carcinoma Hep3B cells
-
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
-
transient replication of HPV-18 mutant ori plasmids with gene-delivered chimeric mutant AZP-SNase, overexpression of recombinant mutant chimeric enzyme, hybrid nuclease AZP-SNase, 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
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
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
-
SaeRS is required for expression of the nuc gene
-
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
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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
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D790E
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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
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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
F34A
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site-directed mutagenesis
G79S/H124LC80-C116
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effects on the stability and conformation of the folded protein
H124LC77-C118
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effects on the stability and conformation of the folded protein
H124LC79-C118
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effects on the stability and conformation of the folded protein
H124LC80-C116
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effects on the stability and conformation of the folded protein
I92A
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site-directed mutagenesis, the mutant shows similar global stability like the wild-type enzyme
INS33A34
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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
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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
L103A
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site-directed mutagenesis, the mutant shows similar global stability like the wild-type enzyme
L125A
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site-directed mutagenesis, the mutant shows similar global stability like the wild-type enzyme
L25A
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site-directed mutagenesis
L36A
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site-directed mutagenesis
L38A
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site-directed mutagenesis
P117G,/H124L/S128A
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site-directed mutagenesis, a highly stable triple mutant
P117G/H124L/S128A
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site-directed mutagenesis
P11A/P31A/P42A/P47T/P56A/P117G
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proline free mutant, conformationally different from wild type protein, 1.4% of wild type activity
P47G/P117G/H124L/W140H
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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
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designed for the insertion of a cysteine reactive label
T62P
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
V23A
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site-directed mutagenesis
V66A
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site-directed mutagenesis, the mutant shows similar global stability like the wild-type enzyme
V66F/P117G,/H124L/S128A
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site-directed mutagenesis of the highly stable triple mutant P117G,/H124L/S128A, thermodynamic stability during guanidine hydrochloride denaturation of mutants is compared
V66G/P117G,/H124L/S128A
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site-directed mutagenesis of the highly stable triple mutant P117G,/H124L/S128A, thermodynamic stability during guanidine hydrochloride denaturation of mutants is compared
V66N/P117G,/H124L/S128A
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site-directed mutagenesis of the highly stable triple mutant P117G,/H124L/S128A, thermodynamic stability during guanidine hydrochloride denaturation of mutants is compared
V66Q/P117G,/H124L/S128A
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site-directed mutagenesis of the highly stable triple mutant P117G,/H124L/S128A, thermodynamic stability during guanidine hydrochloride denaturation of mutants is compared
V66S/P117G,/H124L/S128A
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site-directed mutagenesis of the highly stable triple mutant P117G,/H124L/S128A, thermodynamic stability during guanidine hydrochloride denaturation of mutants is compared
V66T/P117G,/H124L/S128A
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site-directed mutagenesis of the highly stable triple mutant P117G,/H124L/S128A, thermodynamic stability during guanidine hydrochloride denaturation of mutants is compared
V66Y/P117G,/H124L/S128A
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site-directed mutagenesis of the highly stable triple mutant P117G,/H124L/S128A, thermodynamic stability during guanidine hydrochloride denaturation of mutants is compared
V74A
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site-directed mutagenesis
A128S
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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
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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
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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
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Tm value for mutant enzyme is 50.53C, compared to 50.98C for wild-type enzyme
D143K
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charge reversal
D143N
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charge reversal
D146G
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Tm value for mutant enzyme is 50.99C, compared to 50.98C for wild-type enzyme
D19G
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Tm value for mutant enzyme is 52.06C, compared to 50.98C for wild-type enzyme
D19K
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charge reversal
D19L
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charge reversal
D21G
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Tm value for mutant enzyme is 53.74C, compared to 50.98C for wild-type enzyme
D21K
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charge reversal
D21N
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charge reversal
D21N/T33V/T41I/S59A/P117G/A128A
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hyperstable engineered form of staphylococcal nuclease (SNase)
D40G
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Tm value for mutant enzyme is 50.44C, compared to 50.98C for wild-type enzyme
D77A
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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
D77K
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charge reversal
D83G
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Tm value for mutant enzyme is 37.21C, compared to 50.98C for wild-type enzyme
D95G
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Tm value for mutant enzyme is 37.38C, compared to 50.98C for wild-type enzyme
DELTA1-139
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mutant lacks tertiary structure, fluorescence of the mutant is much lower than that of the wild-type enzyme
DELTA1-141
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intact tertiary conformation, melting point is nearly identical to wild-type enzyme
E101A
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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
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Tm value for mutant enzyme is 43.04C, compared to 50.98C for wild-type enzyme
E10G
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Tm value for mutant enzyme is 43.8C, compared to 50.98C for wild-type enzyme
E10K
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charge reversal
E10Q
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charge reversal
E122G
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Tm value for mutant enzyme is 44.12C, compared to 50.98C for wild-type enzyme
E122K
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charge reversal
E122Q
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charge reversal
E129G
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Tm value for mutant enzyme is 34.59C, compared to 50.98C for wild-type enzyme
E135G
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Tm value for mutant enzyme is 44.54C, compared to 50.98C for wild-type enzyme
E135K
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charge reversal
E135Q
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charge neutralization
E142G
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Tm value for mutant enzyme is 49.41C, compared to 50.98C for wild-type enzyme
E43G
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Tm value for mutant enzyme is 54.99C, compared to 50.98C for wild-type enzyme
E52G
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Tm value for mutant enzyme is 52.1C, compared to 50.98C for wild-type enzyme
E57G
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Tm value for mutant enzyme is 46.6C, compared to 50.98C for wild-type enzyme
E57K
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charge reversal
E57Q
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charge reversal
E67G
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Tm value for mutant enzyme is 46.53C, compared to 50.98C for wild-type enzyme
E67Q
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charge reversal
E73A
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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/D77G
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loss of thermal stabilty of 47% relative to the wild-type protein
E73G/E75G
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loss of thermal stabilty of 59% relative to the wild-type protein
E73G/E75G/D77G
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loss of thermal stabilty of 65% relative to the wild-type protein
E73K
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charge reversal
E73Q
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charge reversal
E75A
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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
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Tm value for mutant enzyme is 36.99C, compared to 50.98C for wild-type enzyme
E75G/D77G
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loss of thermal stabilty of 58% relative to the wild-type protein
E75K
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charge reversal
F34W/W140F
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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
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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
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mutation causes decrease in thermal stability
G20A
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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
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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
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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
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hyperstable, acid-resistant mutant form of enzyme known as delta+PHS
G79S
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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
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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
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charge reversal
H124L
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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
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charge neutralization
I92E
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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
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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
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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
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charge reversal
K127Q
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charge neutralization
K133A
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intact tertiary conformation, melting point is 4.6C lower than that of the wild-type enzyme
K133Q
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charge neutralization
K28C/H124C
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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
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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
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charge reversal
K28Q
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charge neutralization
K48E
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charge reversal
K48Q
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charge neutralization
K63E
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charge reversal
K63Q
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charge neutralization
K64E
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charge reversal
K64Q
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charge neutralization
K70E
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charge reversal
K70Q
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charge neutralization
K78E
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charge reversal
K78Q
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charge neutralization
K84E
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charge reversal
K84Q
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charge neutralization
K97E
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charge reversal
K97Q
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charge neutralization
K9A
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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)
L25A
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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
S28C
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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
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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
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production by site-directed mutagenesis, pka value shifts to 8.95 and, after chemical denaturation, to 8.73
V66E/P117G/H124L/S128A
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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
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production by site-directed mutagenesis, pka value shifts to 9.07 and, after chemical denaturation, to 8.80
V66K/P117G/H124L/S128A
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production by site-directed mutagenesis, pka value shifts to 6.35
V66K/P117G/H124L/S128A/G50F/V51N/DELTA44-49
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production by site-directed mutagenesis, pka value shifts to 5.63 and, after chemical denaturation, to 5.83
V66L
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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
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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
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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
W140C
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DNA hydrolysis activity is 75% of wild-type activity
W140D
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DNA hydrolysis activity is 65% of wild-type activity
W140E
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DNA hydrolysis activity is 65% of wild-type activity
W140G
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DNA hydrolysis activity is 75% of wild-type activity
W140I
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DNA hydrolysis activity is 70% of wild-type activity
W140K
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DNA hydrolysis activity is 70% of wild-type activity
W140M
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DNA hydrolysis activity is 70% of wild-type activity
W140N
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DNA hydrolysis activity is 75% of wild-type activity
W140P
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DNA hydrolysis activity is 55% of wild-type activity
W140Q
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DNA hydrolysis activity is 75% of wild-type activity
W140R
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DNA hydrolysis activity is 75% of wild-type activity
W140S
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DNA hydrolysis activity is 75% of wild-type activity
W140T
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DNA hydrolysis activity is 75% of wild-type activity
W140V
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DNA hydrolysis activity is 75% of wild-type activity
Y54C/I139C
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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
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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
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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
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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
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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
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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
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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
additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
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)
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effect of denaturants at low concentrations on very unstable mutant enzyme forms
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high concentration of urea and guanidinium chloride denaturation is completely reversible at pH 3.5-3.7, 55-65C
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refolding of acid-unfolded enzyme induced anions
-
reversible thermal denaturation
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
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
drug development
medicine
molecular biology
Show AA Sequence (287 entries)
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