Information on EC 3.4.21.81 - Streptogrisin B

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The expected taxonomic range for this enzyme is: Streptomyces griseus

EC NUMBER
COMMENTARY hide
3.4.21.81
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RECOMMENDED NAME
GeneOntology No.
Streptogrisin B
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Hydrolysis of proteins with trypsin-like specificity
show the reaction diagram
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-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
CAS REGISTRY NUMBER
COMMENTARY hide
55071-87-9
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
3 acetyl-Pro-Ala-Phe-Phe-NH2 + H2O
acetyl-Pro-Ala-Phe-Phe + NH3 + acetyl-Pro-Ala + Phe-Phe-NH2 + acetyl-Pro-Ala-Phe + Phe-NH2
show the reaction diagram
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hydrolyzed at the Ala-Phe, the Phe-Phe and the Phe-NH2 bond
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-
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acetyl-Ala-Ala-Tyr-4-methylcoumarin 7-amide + H2O
acetyl-Ala-Ala-Tyr + 7-amino-4-methylcoumarin
show the reaction diagram
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-
-
-
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Acetyl-Pro-Ala-Ala-Phe-NH2 + H2O
?
show the reaction diagram
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split at the amide bond as well as at the Ala-Ala bond
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-
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Acetyl-Pro-Ala-Gly-Phe-NH2 + H2O
?
show the reaction diagram
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split at the amide bond as well as at the Ala-Gly bond
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-
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Acetyl-Pro-Ala-Leu-Phe-NH2 + H2O
Acetyl-Pro-Ala-Leu-Phe-OH + NH3
show the reaction diagram
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hydrolyzed at the amide bond only
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-
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Acetyl-Pro-Ala-Pro-Phe-NH2 + H2O
Acetyl-Pro-Ala-Pro-Phe-OH + NH3
show the reaction diagram
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hydrolyzed at the amide bond only
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-
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N-Acetyl-L-Pro-L-Leu-p-nitroanilide + H2O
?
show the reaction diagram
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-
-
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OMSVP3 + H2O
?
show the reaction diagram
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ovomucoid third domain from silver pheasant, potent inhibitor of chymotrypsin, subtilisin, and elastase
cleavage between residues Met18-Glu19
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?
OMSVP3 mutant P14C/N39C + H2O
?
show the reaction diagram
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disulfide variant of the ovomucoid third domain from silver pheasant carrying an engineered Cys14-Cys39 bond near the reactive site
cleavage between residues Met18-Glu19. P14C/N39C can be selectively cleaved by Streptomyces griseus protease B at the reactive site of OMSVP3 to form a reactive site modified inhibitor. The conversion rate of P14C/N39C is much faster than that for wild type under any pH condition. The reactive site modified form of P14C/N39C is thermodynamically more stable than the intact one
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?
Sc-AAPF-p-nitroanilide + H2O
?
show the reaction diagram
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-
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?
Sc-AAPF-SBn + FAASF-NH2
?
show the reaction diagram
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ligation to nonapeptide by mutant S195A/T213L/F228H
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?
Sc-AAPF-SBn + H2O
?
show the reaction diagram
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-
-
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Suc-AAPF-p-nitroanilide + H2O
?
show the reaction diagram
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-
-
?
Suc-AAPF-SBzl + H2O
Suc-AAPF + phenylmethanethiol
show the reaction diagram
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-
-
?
succinyl-Ala-Ala-Pro-4-methylcoumarin 7-amide + H2O
succinyl-Ala-Ala-Pro + 7-amino-4-methylcoumarin
show the reaction diagram
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succinyl-Ala-Ala-Pro-Phe 4-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
show the reaction diagram
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-
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succinyl-Ala-Ala-Pro-X-4-nitroanilide + H2O
?
show the reaction diagram
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-
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?
succinyl-Ala-Ala-Pro-X-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
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?
succinyl-L-Ala-L-Ala-L-Pro-L-Asp-4-nitroanilide + H2O
succinyl-L-Ala-L-Ala-L-Pro-L-Asp + 4-nitroaniline
show the reaction diagram
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extremely poor substrate at pH 8 but becomes a moderately good substrate at pH 5.5
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-
?
succinyl-L-Ala-L-Ala-L-Pro-L-Leu-4-nitroanilide + H2O
succinyl-L-Ala-L-Ala-L-Pro-L-Leu + 4-nitroaniline
show the reaction diagram
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optimum at pH 7.5-8.0
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?
succinyl-L-Ala-L-Ala-L-Pro-L-Lys-4-nitroanilide + H2O
succinyl-L-Ala-L-Ala-L-Pro-L-Lys + 4-nitroaniline
show the reaction diagram
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optimum at pH 10.0
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?
additional information
?
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Bovine pancreatic trypsin inhibitor
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Kunitz
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N-tert-Butyloxycarbonyl-glycyl-L-leucyl-L-phenylalanine chloromethyl ketone
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N-tert-Butyloxycarbonyl-L-alanylglycyl-L-phenylalanine chloromethyl ketone
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protein inhibitor S13D14Y15-OMTKY3
a OMTKY3 mutant variant, recombinantly produced in Escherichia coli strain RV308 and purified, strong inhibition, overview
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protein inhibitor S13D14Y15G18I19K21-OMTKY3
a OMTKY3 mutant variant, recombinantly produced in Escherichia coli strain RV308 and purified, strong inhibition, overview. S13D14Y15G18I19K21 OMTKY3 inhibits Streptomyces griseus protease B 10times more strongly than Streptomyces griseus protease A, EC 3.4.21.80
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Soybean trypsin inhibitor
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Squash inhibitor
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Streptomyces subtilisin inhibitor
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Third domain of the ovomucoid inhibitor from turkey
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Turkey ovomucoid third domain
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OMTKY3, inhibitor for serine peptidases
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additional information
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.15
Acetyl-Ala-Ala-Tyr-4-methylcoumarin 7-amide
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1.4
Acetyl-Pro-Ala-Leu-Phe-NH2
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2.4
Acetyl-Pro-Ala-Pro-Phe-NH2
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0.062 - 0.5
Suc-AAPF-p-nitroanilide
0.101
succinyl-Ala-Ala-Pro-Phe 4--nitroanilide
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0.098
Succinyl-Ala-Ala-Pro-Phe-4-methylcoumarin 7-amide
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additional information
additional information
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.04
Acetyl-Ala-Ala-Tyr-4-methylcoumarin 7-amide
Streptomyces griseus
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7.1
Acetyl-Pro-Ala-Leu-Phe-NH2
Streptomyces griseus
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6.7
Acetyl-Pro-Ala-Pro-Phe-NH2
Streptomyces griseus
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0.066 - 1884
Suc-AAPF-p-nitroanilide
153
succinyl-Ala-Ala-Pro-Phe 4-nitroanilide
Streptomyces griseus
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3.3
Succinyl-Ala-Ala-Pro-Phe-4-methylcoumarin 7-amide
Streptomyces griseus
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additional information
additional information
Streptomyces griseus
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
740 - 10700
succinyl-L-Ala-L-Ala-L-Pro-L-Asp-4-nitroanilide
168017
28200 - 25000000
succinyl-L-Ala-L-Ala-L-Pro-L-Leu-4-nitroanilide
168018
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 6
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substrate succinyl-L-Ala-L-Ala-L-Pro-L-Asp-4-nitroanilide
6
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hydrolysis reaction
7.5 - 8
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substrate succinyl-L-Ala-L-Ala-L-Pro-L-Leu-4-nitroanilide
8.8 - 9.2
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ligation reaction of mutant S195A/T213L/F228H
10
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substrate succinyl-L-Ala-L-Ala-L-Pro-L-Lys-4-nitroanilide
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
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assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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ratio of ligation to hydrolysis by mutant S195A/T213L/F228H decreases with increase in temperature from 30C to 60C
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
18410
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Streptomyces griseus, amino acid sequence
18412
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1 * 18412, Streptomyces griseus, amino acid sequence
additional information
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amino acid sequence of cysteic acid peptides from peptic digest
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
additional information
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enzyme folds to native state that is thermodynamically marginally stable, apparent stability arises kinetically from unfolding free energy barriers. Enzyme has effective pro-regions to facilitate folding
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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enzyme has effective pro-regions to facilitate folding
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
bound to wild type OMTKY3 and to the Ala32I variant, by the hanging-drop vapor-diffusion method, SGPB and OMTKY3 Ala32I forms a 1 : 2 complex in the crystal
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crystal structure in complex with turkey ovomucoid thir domain backbone variants OMTKY3-Pro18I and OMTKY3-psi[COO]-Leu18I
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crystal structure of protease B with tripeptide chloromethyl ketone inhibitors
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crystal structures of Leu18, Ala18, and Gly18 variants of turkey ovomucoid inhibitor third domain complexed with proteinase B
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structure at 2.8 A resolution
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structure of the complex of protease B with the third domain of the ovomucoid inhibitor from turkey at 1.8 A resolution
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structure of the complex of proteinase B and polypeptide chymotrypsin inhibitor-1 from Russet Burbank potato tubers at 2.1 A resolution
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native crystal structures at pH 4.2 at a resolution of 1.18 A, and at pH 7.3 at a resolution of 1.23 A. At pH 4.2 the enzyme is assigned as a tetrapeptide, Asp-Ala-Ile-Tyr, whereas at pH 7.3 it is assigned as a tyrosine molecule and a leucine molecule existing at equal occupancies in both of the SGPB molecules in the asymmetric unit
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37 - 45
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mutant S195A, completely inactivated in less than 1 min at 45C, less than 20 min at 37C
additional information
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study of temperature-dependences of the folding and unfolding kinetics without enzyme pro region
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
Stable in presence of 6.0 M guanidinium chloride
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ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
guanidine-HCl
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8 M, extremely stable
urea
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6 M, extremely stable
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
further purification of a commercial preparation
mutants S195A and S195G
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purified from a commercial preparation of Pronase
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purified from Pronase
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
active-site variant genes clones in the Escherichia coli/Bacillus subtilis shuttle vector pEB-11 and expressed in Bacillus subtilis DB104
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gene structure
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genes constructed which encodes deletions at the amino-terminal end of the propeptide, deletions to the 76-amino-acid propeptide of SGPB cause an incremental loss in the production of the mature enzyme
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
S195A
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active-site variant of protease B with peptide-ligation activity
S195A/T213L/F228H
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i.e. streptoligase, catalyzes peptide ligation efficiently. Ligation proceeds via an acyl-enzyme intermediate involving H57. Mutant exhibits half-life for unfolding of 16.3 min at 55C in the absence of stabilizing substrates
S195C
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active-site variant of protease B with peptide-ligation activity
S195G
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active-site variant of protease B with peptide-ligation activity
S195G/H57N
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active-site variant of protease B with peptide-ligation activity
additional information
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introduction of site-specific disulfide bonds into the flexible N-terminal loop of natural Kazal-type inhibitors to characterize the thermodynamics of the reactive site peptide bond hydrolysis. Mutant P14C/N39C is a disulfide variant of the ovomucoid third domain from silver pheasant carrying an engineered Cys14Cys39 bond near the reactive site. The conversion rate of P14C/N39C by streptogrisin B is much faster than that for wild type under any pH condition
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
study of temperature-dependencies of the folding and unfolding kinetics without enzyme pro region. Enzyme shows a maximal unfolding cooperativity and slow rate of global unfolding
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