Information on EC 3.4.22.25 - glycyl endopeptidase

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The expected taxonomic range for this enzyme is: Carica papaya

EC NUMBER
COMMENTARY
3.4.22.25
-
RECOMMENDED NAME
GeneOntology No.
glycyl endopeptidase
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
preferential cleavage: Gly-/-, in proteins and small molecule substrates
show the reaction diagram
-
-
-
-
preferential cleavage: Gly-/-, in proteins and small molecule substrates
show the reaction diagram
specific for Gly at posotion P1 due to an exchange conserved residues Gly23 to Glu and Gly65 to Arg, 2 free thiol functions of cysteinyl residues are essential for the catalytic competence of the enzyme
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
-
-
hydrolysis of peptide bond
-
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Chymopapain
-
-
-
-
Chymopapain M
-
-
-
-
Chymopapain M
-
-
glycine endopeptidase
-
-
Glycyl endopeptidase
-
-
-
-
Papaya peptidase B
-
-
-
-
Papaya peptidase B
-
-
Papaya proteinase 4
-
-
-
-
Papaya proteinase IV
-
-
-
-
Papaya proteinase IV
-
-
PPIV
-
-
-
-
Proteinase, glycine-specific
-
-
-
-
Glycyl endopeptidase
-
-
additional information
-
the enzyme belongs to the C1A peptidase family
CAS REGISTRY NUMBER
COMMENTARY
149719-24-4
-
92228-52-9
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
from commercial chymopapain
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
aggrecan + H2O
?
show the reaction diagram
-
-
-
-
-
azocasein + H2O
trichloroacetic acid-soluble peptides from azocasein
show the reaction diagram
-
10% of the activity of papain
-
-
benzyloxycarbonyl-Gly-Leu-NH + H2O
L-leucine amide + N-benzyloxycarbonyl-glycine
show the reaction diagram
P05994
-
-
-
?
benzyloxycarbonyl-Gly-Phe-NH + H2O
L-phenylalanine amide + N-benzyloxycarbonyl-glycine
show the reaction diagram
P05994
-
-
-
?
benzyloxycarbonyl-Gly-Tyr-NH + H2O
L-tyrosine amide + N-benzyloxycarbonyl-glycine
show the reaction diagram
P05994
-
-
-
?
benzyloxycarbonyl-Gly-Tyr-Oet + H2O
L-tyrosine ethyl ester + N-benzyloxycarbonyl-glycine
show the reaction diagram
P05994
-
-
-
?
casein + H2O
trichloroacetic acid-soluble peptides derived from casein
show the reaction diagram
-
10% of the activity of papain
-
-
Chicken cystatin + H2O
?
show the reaction diagram
-
-
-
-
-
Chicken cystatin + H2O
?
show the reaction diagram
-
-
-
-
?
Gly-4-nitroanilide + H2O
glycine + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
Hemoglobin + H2O
Trichloroacetic acid-soluble peptides from haemoglobin
show the reaction diagram
-
-
-
-
hide powder azure + H2O
?
show the reaction diagram
-
-
-
-
-
Human cystatin C + H2O
?
show the reaction diagram
-
-
-
-
-
N-Acetyl-L-Phe-Gly 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
N-Benzyloxycarbonyl-Gly 4-nitrophenyl ester + H2O
?
show the reaction diagram
-
-
-
-
-
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
tert-Butyloxycarbonyl-Ala-Ala-Ala 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
-
-
-
-
tert-Butyloxycarbonyl-Ala-Ala-Gly 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
-
-
-
-
tert-Butyloxycarbonyl-Ala-Ala-Gly 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
tert-butyloxycarbonyl-Ala-Ala-Gly-p-nitroanilide + H2O
tert-butyloxycarbonyl-Ala-Ala-Gly + p-nitroaniline
show the reaction diagram
P05994
-
-
-
?
Trypsin + H2O
Hydrolyzed trypsin
show the reaction diagram
-
16 cleavage points of which 13 are glycyl bonds
-
-
Human cystatin C + H2O
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
synthetic substrates of other papaya proteinases
-
-
-
additional information
?
-
-
absolute specificity for substrates with glycine at the P1 position (structural origin of the unusual specificity)
-
-
-
additional information
?
-
-
not: cystatin A
-
-
-
additional information
?
-
-
the recombinant proregion of the enzyme can inhibit the cysteine protease of larvae of the Colorado beetle, Leptinotarsa decemlineata, analoguesly to specific inhibitor oryzacystatin I, inhibitory effect is affected by temperature
-
?
additional information
?
-
-
the enzyme is useful for the plant defense system, substrate specificity with preference for glycyl residues, structure-function relationship, overview, hydrolyzes glycine esters but not Bz-Arg-4-nitroanilide
-
-
-
additional information
?
-
P05994
glycyl endopeptidase fails to hydrolyse DL-benzyl-Arg-p-nitroanilide
-
-
-
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
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
additional information
?
-
-
the recombinant proregion of the enzyme can inhibit the cysteine protease of larvae of the Colorado beetle, Leptinotarsa decemlineata, analoguesly to specific inhibitor oryzacystatin I, inhibitory effect is affected by temperature
-
?
additional information
?
-
-
the enzyme is useful for the plant defense system
-
-
-
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Alpha-macroglobulin
-
-
-
E64
-
inactivation
iodoacetamide
-
low rate of inactivation
iodoacetamide
-
slow inactivation
iodoacetate
-
low rate of inactivation
iodoacetate
-
slow inactivation
papaya cystatin
-
-
-
Peptide aldehydes
-
despite the presence of bulky sidechains in P1
Peptidyl diazomethanes
-
containing glycine in P1 and a hydrophobic side-chain in P2
Peptidyl diazomethanes
-
-
Peptidyl diazomethanes
-
dependent on the substrate
Pro-peptide
-
the enzyme is inhibited by its own pro region after cleavage, also pro-peptide of other C1 peptidase family enzymes can be inhibitory
-
L-3-carboxy-2,3-epoxypropionylleucylamido(4-guanidino)butane
-
-
additional information
-
human cystatin A; human cystatin C; not: chicken cystatin
-
additional information
-
not: chicken cystatin
-
additional information
-
cystatin family of cysteine proteinase inhibitors
-
additional information
-
not: chicken cystatin (structural origin of the lack of inhibition)
-
additional information
-
no inhibition by most cystatins
-
additional information
P05994
not inhibited by cystatin
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
-
the enzyme depends on a thiolate ion for activity
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.08
-
tert-butyloxycarbonyl-Ala-Ala-Ala 4-methylcoumarin 7-amide
-
-
0.16
-
tert-butyloxycarbonyl-Ala-Ala-Gly 4-methylcoumarin 7-amide
-
-
5.2
-
tert-butyloxycarbonyl-Ala-Ala-Gly 4-nitroanilide
-
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.08
-
tert-butyloxycarbonyl-Ala-Ala-Ala 4-methylcoumarin 7-amide
-
-
5
-
tert-butyloxycarbonyl-Ala-Ala-Gly 4-methylcoumarin 7-amide
-
-
22
-
tert-butyloxycarbonyl-Ala-Ala-Gly 4-nitroanilide
-
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0000034
-
papapya cystatin
-
-
-
0.00086
-
Pro-peptide
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7
7.5
-
tert-butyloxycarbonyl-Ala-Ala-Gly 4-nitroanilide
7
-
-
azocasein
7
-
-
about
8
-
-
assay at
additional information
-
-
pI: 10.6-10.7
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3
9.5
-
3.0: 15% of maximal activity, 9.5: about 75% of maximal activity, tert-butyloxycarbonyl-Ala-Ala-Gly 4-nitroanilide
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
23310
-
-
Carica papaya, calculation from amino acid sequence
24000
-
-
Carica papaya, sedimentation analysis, meniscus depletion method
additional information
-
-
three-dimensional structure
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
monomer
-
1 * 23313
additional information
-
three-dimensional structure comparison to other latex peptidase
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
no glycoprotein
-
no site of potential glycosylation detected
no glycoprotein
-
mature enzyme
proteolytic modification
-
the enzyme is synthesized as preproenzyme
additional information
-
enzyme is synthesized as inactive proenzyme, and rapidly converted to the active from within 2 min after wounding of the plant
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
crystal structure
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
the enzyme undergoes a conformational transition at low pH and 37°C, that instantaneously and irreversibly converts the native form into molten globules, which are unstable and rapidly degraded by pepsin
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
recombinant proregion, 60 min, stable
additional information
-
-
factors effecting thermostability
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ammonium sulfate precipitation
P05994
different methods, overview
-
native enzyme from latex by ion exchange and adsorption chromatography, and gel filtration
-
several ligands possible for affinity chromatography are presented, fractionation on hydrophobic and cation-exchange supports, overview
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression of the pro-region of papaya proteinase IV in Escherichia coli
-
expression of the proregion of the enzyme in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
G27E
-
site-directed mutagenesis, high reduced activity with substrate having an Arg at P1 position
G73R
-
site-directed mutagenesis, high reduced activity with substrate having an Arg at P1 position, decreased affinity for cystatin C
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
biotechnology
-
plant protease proregions have a potential as regulators of cysteine proteinases in biotechnological systems and to target proteases of pests
nutrition
-
the enzyme needs to be protected from acid denaturation and proteolysis in the gut after oral administration to be effective as cysteine protease