Information on EC 3.4.22.10 - streptopain

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The expected taxonomic range for this enzyme is: Streptococcus

EC NUMBER
COMMENTARY hide
3.4.22.10
-
RECOMMENDED NAME
GeneOntology No.
streptopain
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
preferential cleavage with hydrophobic residues at P2, P1 and P1'
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
-
endopeptidase
-
CAS REGISTRY NUMBER
COMMENTARY hide
9025-51-8
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain A-20
-
-
Manually annotated by BRENDA team
serotype M1
-
-
Manually annotated by BRENDA team
strain B220
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
precursor; strain HSC5
UniProt
Manually annotated by BRENDA team
serotype M3
-
-
Manually annotated by BRENDA team
serotype M1
-
-
Manually annotated by BRENDA team
Streptococcus sp. strainT1BRB
strainT1BRB
-
-
Manually annotated by BRENDA team
strain T10BW
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
-
wild type strain efficiently translocates across the epithelial monolayer along with cleavage of occludin and E-cadherin. Deletion of the speB gene compromises those activities. Destabilization of the junctional proteins is relieved in cells infected with the speB mutant, as compared with those infected with the wild type
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
3-aminobenzoyl-Ala-Ala-Ile-Lys-Ala-Gly-Ala-Arg + H2O
?
show the reaction diagram
3-aminobenzoyl-Ala-Glu-Ile-Lys-Gln-Pro-Val-Val + H2O
?
show the reaction diagram
-
-
-
-
?
3-aminobenzoyl-Asp-Lys-Val-Asn-Leu-Gly-Gly-Glu + H2O
?
show the reaction diagram
3-aminobenzoyl-Glu-Gln-Ile-Lys-Glu-Asn-Lys-Lys + H2O
?
show the reaction diagram
3-aminobenzoyl-Thr-Thr-Thr-Ala-Gly-Thr-Ala-Glu + H2O
?
show the reaction diagram
acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin + H2O
?
show the reaction diagram
alpha1-antitrypsin + H2O
?
show the reaction diagram
-
-
-
-
?
azocasein + H2O
?
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Ala + H2O
?
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Glu-p-nitrophenyl ester + H2O
?
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Glu-Phe + H2O
?
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Gly-Gly-p-nitrophenyl ester + H2O
?
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Gly-p-nitrophenyl ester + H2O
benzyloxycarbonyl-Gly + p-nitrophenol
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Nle-p-nitrophenyl ester + H2O
?
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Nle-Phe + H2O
?
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Val-Arg-4-methylcoumarinyl-7-amide hydrochloride + H2O
?
show the reaction diagram
Bovine serum albumin + H2O
?
show the reaction diagram
C5a peptidase + H2O
?
show the reaction diagram
-
cleavage sites are APA-K, AVI-D, SGTS, and C-terminally
-
-
?
casein + H2O
?
show the reaction diagram
cinnamoyl-Gly-p-nitrophenyl ester + H2O
cinnamoyl-Gly + 4-nitrophenol
show the reaction diagram
-
-
-
-
?
complement component C3 + H2O
?
show the reaction diagram
-
-
-
-
?
complement factor C3b + H2O
?
show the reaction diagram
-
cleavage sites are RTL-D, NHK-L, LAR-S, and VEL-I
-
-
?
D-Ile-Pro-Arg-p-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
decorin + H2O
?
show the reaction diagram
-
-
-
-
?
E-cadherin + H2O
?
show the reaction diagram
-
-
-
-
?
EndoS + H2O
?
show the reaction diagram
-
cleavage site is VML-K
-
-
?
Fba + H2O
?
show the reaction diagram
Fba protein + H2O
?
show the reaction diagram
-
-
-
-
?
Fibrinogen + H2O
?
show the reaction diagram
Fibronectin + H2O
?
show the reaction diagram
-
-
-
-
?
H-kininogen + H2O
?
show the reaction diagram
-
cleavage sites are LMK-R and PFR-S
-
-
?
H-kininogen + H2O
biologically active kinins
show the reaction diagram
-
-
-
?
human CXCL-1 + H2O
?
show the reaction diagram
human CXCL-2 + H2O
?
show the reaction diagram
human CXCL-6 + H2O
?
show the reaction diagram
human CXCL-8 + H2O
?
show the reaction diagram
human CXCL10 + H2O
?
show the reaction diagram
-
-
-
-
?
Human fibrinogen + H2O
?
show the reaction diagram
human interleukin 1beta precursor + H2O
human interleukin 1beta + fragment
show the reaction diagram
-
-
-
-
?
IgA + H2O
?
show the reaction diagram
IgD + H2O
?
show the reaction diagram
-
degradation of the carboxy-terminal part of the heavy chain
-
?
IgE + H2O
?
show the reaction diagram
-
degradation of the carboxy-terminal part of the heavy chain
-
?
IgG + H2O
?
show the reaction diagram
IgG + H2O
Fab fragment + Gc fragments
show the reaction diagram
-
the enzyme cleaves IgG in the flexible hinge region of the IgG heavy chain, generating two Fab fragments and Fc fragment. SpeB cleaves the heavy chain at a defined site between glycine residues 236 and 237
-
?
IgM + H2O
?
show the reaction diagram
immunoglobulin A + H2O
?
show the reaction diagram
-
cleavage sites are the heavy chains
-
-
?
immunoglobulin D + H2O
?
show the reaction diagram
-
cleavage sites are the heavy chains
-
-
?
immunoglobulin E + H2O
?
show the reaction diagram
-
cleavage sites are the heavy chains
-
-
?
immunoglobulin G + H2O
?
show the reaction diagram
-
cleavage site is LLG-G in gamma-chain
-
-
?
immunoglobulin G + H2O
F(ab')2 + 1/2Fc
show the reaction diagram
-
non-immune binding of IgG to the bacterial surface is followed by the proteolytic cleavage of the antibody by the IgG-endopeptidase IdeS. IdeS generated 1/2Fc fragments do not compete efficiently with intact IgG in binding to the bacterial surface and rapid dissociation of 1/2Fc allows binding of new IgG. A correlated binding and proteolytic cleavage of IgG increases the probability that the bacteria can resist specific IgG, despite the presence of a large excess of non-specific IgG in the circulation. As a consequence of IdeS activity, circulating 1/2Fc fragments are generated. These 1/2Fc fragments are shown to be biological active by acting as priming agents for polymorphonuclear leucocytes
-
-
?
immunoglobulin G + H2O
F(ab')2 fragment + 1/2 Fc fragments
show the reaction diagram
-
IdeS cleaves IgG by hydrolyzing the peptide bond between two glycine residues in the hinge region of IgG, generating one F(ab')2 fragment and two 1/2 Fc fragments
-
-
?
immunoglobulin M + H2O
?
show the reaction diagram
-
cleavage sites are the heavy chains
-
-
?
insulin + H2O
?
show the reaction diagram
-
reduced carboxymethylated phenylalanine chain, one of the most rapidly hydrolyzed Phe-Tyr linkages is in the Phe-Phe-Tyr sequence
-
-
?
insulin beta chain + H2O
?
show the reaction diagram
-
reduced and carboxylated substrate, cleavage of the linkage Phe25-Tyr26
-
-
?
interleukin-1beta precursor + H2O
interleukin-1beta + interleukin-1beta propeptide
show the reaction diagram
Laminin + H2O
?
show the reaction diagram
-
-
-
-
?
M protein + H2O
?
show the reaction diagram
-
-
-
-
?
mouse CXCL-1 + H2O
?
show the reaction diagram
mouse CXCL-2 + H2O
?
show the reaction diagram
-
the cleavage site is between lysine residues K65 and K66
-
-
?
N-benzoyl-Gly-p-nitrophenyl ester + H2O
N-benzoyl-Gly + 4-nitrophenol
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-Ala-p-nitrophenyl ester + H2O
N-benzyloxycarbonyl-Ala + 4-nitrophenol
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-Gly + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-Gly-p-nitrophenyl ester + H2O
N-benzyloxycarbonyl-Gly + 4-nitrophenol
show the reaction diagram
Nalpha-benzyloxycarbonyl-Lys-p-nitrophenyl ester + H2O
?
show the reaction diagram
-
-
-
-
?
Nalpha-benzyloxycarbonyl-Lys-Phe + H2O
?
show the reaction diagram
-
-
-
-
?
Nalpha-benzyloxycarbonyl-Nepsilon-t-butyloxycarbonyl-Lys-Phe + H2O
?
show the reaction diagram
-
-
-
-
?
Nalpha-benzyloxycarbonyl-Nepsilon-tosyl-Lys-p-nitrophenyl ester + H2O
Nalpha-benzyloxycarbonyl-Nepsilon-tosyl-Lys + p-nitrophenol
show the reaction diagram
-
-
-
-
?
Nalpha-benzyloxycarbonyl-Phe-Phe + H2O
?
show the reaction diagram
-
-
-
-
?
Nalpha-benzyloxycarbonyl-Phe-Tyr + H2O
?
show the reaction diagram
-
-
-
-
?
Nepsilon-benzyloxycarbonyl-Lys-p-nitrophenyl ester + H2O
Nepsilon-benzyloxycarbonyl-Lys + 4-nitrophenol
show the reaction diagram
-
-
-
-
?
occludin + H2O
?
show the reaction diagram
-
-
-
-
?
p-nitrobenzyloxycarbonyl-Gly-p-nitrophenyl ester + H2O
p-nitrobenzyloxycarbonyl-Gly + 4-nitrophenol
show the reaction diagram
-
-
-
-
?
p-nitrophenyl-benzyloxycarbonyl-L-Ala + H2O
?
show the reaction diagram
-
-
-
-
?
plasminogen + H2O
?
show the reaction diagram
-
-
-
-
?
pro-interleukin-1beta + H2O
interleukin-1beta + ?
show the reaction diagram
-
cleavage site is YVH-D
-
-
?
properdin + H2O
?
show the reaction diagram
proSPE B C47S + H2O
?
show the reaction diagram
-
the proSPE B C47S mutant is also be used as the substrate for activity assay because it does not exhibit any enzyme activity and exists as a 42 kDa zymogen
-
-
?
proSpeB + H2O
SpeB + ?
show the reaction diagram
protein F1 + H2O
?
show the reaction diagram
protein H + H2O
?
show the reaction diagram
-
-
-
-
?
resorufin-labeled casein + H2O
?
show the reaction diagram
-
-
-
-
?
Sda1 + H2O
?
show the reaction diagram
-
-
-
-
?
Slr + H2O
?
show the reaction diagram
-
-
-
-
?
SmeZ + H2O
?
show the reaction diagram
-
-
-
-
?
SpeBz + H2O
mature SpeB + ?
show the reaction diagram
-
auto-cleavage sites are AIK-A, KVN-L, QIK-E, TYA-G, and EIK-Q
-
-
?
streptococcal mitogenic exotoxin SmeZ + H2O
?
show the reaction diagram
-
cleaves between glutamate and glycine residues
-
-
?
streptococcal mitogenic exotoxin SpeG + H2O
?
show the reaction diagram
-
partial degradation, SpeB cleaves between glutamate and glycine residues
-
-
?
streptococcal mitogenic exotoxin Z + H2O
?
show the reaction diagram
-
-
-
-
?
streptokinase + H2O
?
show the reaction diagram
-
-
-
-
?
streptolysin O + H2O
?
show the reaction diagram
-
-
-
-
?
t-butyloxycarbonyl-Gly-p-nitrophenyl ester + H2O
t-butyloxycarbonyl-Gly + 4-nitrophenol
show the reaction diagram
-
-
-
-
?
tert-butyloxycarbonyl-Ala-p-nitrophenyl ester + H2O
tert-butyloxycarbonyl-Ala + 4-nitrophenol
show the reaction diagram
-
-
-
-
?
urokinase receptor + H2O
?
show the reaction diagram
-
cleavage site is close to the GPI anchor
-
-
?
Vitronectin + 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
alpha1-antitrypsin + H2O
?
show the reaction diagram
-
-
-
-
?
C5a peptidase + H2O
?
show the reaction diagram
-
cleavage sites are APA-K, AVI-D, SGTS, and C-terminally
-
-
?
complement factor C3b + H2O
?
show the reaction diagram
-
cleavage sites are RTL-D, NHK-L, LAR-S, and VEL-I
-
-
?
decorin + H2O
?
show the reaction diagram
-
-
-
-
?
EndoS + H2O
?
show the reaction diagram
-
cleavage site is VML-K
-
-
?
Fba + H2O
?
show the reaction diagram
-
a cell surface factor H- and factor H-like protein 1-binding protein of epithelial human cells, the enzyme inhibits complement regulatory protein factor H- and factor H-like protein 1 binding by proteolysis of the Fba protein, overview
-
-
?
Fba protein + H2O
?
show the reaction diagram
-
-
-
-
?
Fibrinogen + H2O
?
show the reaction diagram
-
cleavage sites are C-terminally in Aalpha chains
-
-
?
Fibronectin + H2O
?
show the reaction diagram
-
-
-
-
?
H-kininogen + H2O
?
show the reaction diagram
-
cleavage sites are LMK-R and PFR-S
-
-
?
human CXCL-1 + H2O
?
show the reaction diagram
human CXCL-2 + H2O
?
show the reaction diagram
human CXCL-6 + H2O
?
show the reaction diagram
human CXCL-8 + H2O
?
show the reaction diagram
human CXCL10 + H2O
?
show the reaction diagram
-
-
-
-
?
Human fibrinogen + H2O
?
show the reaction diagram
-
release and processing, the enzyme abolishes the binding of Streptococcus pyogenes to full-length or 30 kDa fragment of human fibrinogen Fn, kinetics overview
-
-
?
IgG + H2O
?
show the reaction diagram
immunoglobulin A + H2O
?
show the reaction diagram
-
cleavage sites are the heavy chains
-
-
?
immunoglobulin D + H2O
?
show the reaction diagram
-
cleavage sites are the heavy chains
-
-
?
immunoglobulin E + H2O
?
show the reaction diagram
-
cleavage sites are the heavy chains
-
-
?
immunoglobulin G + H2O
?
show the reaction diagram
-
cleavage site is LLG-G in gamma-chain
-
-
?
immunoglobulin M + H2O
?
show the reaction diagram
-
cleavage sites are the heavy chains
-
-
?
Laminin + H2O
?
show the reaction diagram
-
-
-
-
?
M protein + H2O
?
show the reaction diagram
-
-
-
-
?
mouse CXCL-1 + H2O
?
show the reaction diagram
mouse CXCL-2 + H2O
?
show the reaction diagram
-
the cleavage site is between lysine residues K65 and K66
-
-
?
plasminogen + H2O
?
show the reaction diagram
-
-
-
-
?
pro-interleukin-1beta + H2O
interleukin-1beta + ?
show the reaction diagram
-
cleavage site is YVH-D
-
-
?
properdin + H2O
?
show the reaction diagram
protein F1 + H2O
?
show the reaction diagram
protein H + H2O
?
show the reaction diagram
-
-
-
-
?
Sda1 + H2O
?
show the reaction diagram
-
-
-
-
?
Slr + H2O
?
show the reaction diagram
-
-
-
-
?
SmeZ + H2O
?
show the reaction diagram
-
-
-
-
?
SpeBz + H2O
mature SpeB + ?
show the reaction diagram
-
auto-cleavage sites are AIK-A, KVN-L, QIK-E, TYA-G, and EIK-Q
-
-
?
streptococcal mitogenic exotoxin Z + H2O
?
show the reaction diagram
-
-
-
-
?
streptokinase + H2O
?
show the reaction diagram
-
-
-
-
?
streptolysin O + H2O
?
show the reaction diagram
-
-
-
-
?
urokinase receptor + H2O
?
show the reaction diagram
-
cleavage site is close to the GPI anchor
-
-
?
Vitronectin + H2O
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
bisulfite
-
reduction is essential for catalytic activity
cyanide
-
reduction is essential for catalytic activity
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(S)-1-azido-2-tosylamino-hexane
-
moderate inhibitor of IdeS
(S)-1-azido-3-methyl-2-tosylamino-butane
-
moderate inhibitor of IdeS
(S)-1-azido-3-phenyl-2-tosylamino-propane
-
moderate inhibitor of IdeS
(S)-3-tosylamino-1-heptanal
-
strong inhibitory activity
(S)-3-tosylamino-heptane-1-nitrile
-
the compound shows some inhibition of IdeS
(S)-4-methyl-3-tosylamino-1-pentanal
-
strong inhibitory activity
(S)-4-phenyl-3-tosylamino-1-butanal
-
strong inhibitory activity
(S)-7-tert-butoxycarbonylamino-3-tosylamino-1-heptanal
-
strong inhibitory activity
1,3-Dibromoacetone
-
rapid and complete loss of activity
acetonitrile
-
mixed type
acetyl-Ala-Gln-Ile-(S)-2,6-diaminohexanal
-
-
-
benzyloxycarbonyl-Ala-Ala-CHN2
-
-
benzyloxycarbonyl-Ala-Ala-Pro-CHN2
-
-
benzyloxycarbonyl-Ala-Phe-Ala-CHN2
-
-
benzyloxycarbonyl-Asn-Val-Gly-CHN2
-
inhibition of protease activity, no influence on the superantigenic properties of the enzyme
Benzyloxycarbonyl-Gly-Phe
-
competitive against benzyloxycarbonyl-Phe-Tyr
benzyloxycarbonyl-Leu-Val-Gly-CHN2
-
inhibits kininogen degradation
Benzyloxycarbonyl-Phe-Ala-CHN2
-
-
benzyloxycarbonyl-Phe-Gly-CHN2
-
-
benzyloxycarbonyl-Phe-Gly-Phe-CHN2
-
-
Cu2+
-
Cu2+ inhibits the proteolytic activity of mature SpeB
dimethyl sulfoxide
-
competitive
Dinitrofluorobenzene
-
-
-
E64
-
i.e. trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane
Gly-Phe-CHN2
-
-
iodoacetamide
-
-
iodoacetate
-
irreversible inhibitor
iodoacetic acid
-
-
L-trans-epoxysuccinyl-leucylamido(4-guanidino)butane
-
-
methyliodine
-
-
-
N-(N-(L-3-trans-carboxyox-irane-2-carbonyl)-L-leucyl)-agmatine
-
-
-
N-[N-(L-3-transcarboxyirane-2-carbonyl)-L-leucyl]agmatine
-
Nalpha-benzyloxycarbonyl-Phe-Phe
-
strong inhibition of hydrolysis of Nalpha-benzyloxycarbonyl-Phe-Tyr
Sodium tetrathionate
-
-
Spe B pro-peptide
-
specific inhibition of the mature enzyme, secondary structure and SpeB binding analysis, inhibition modeling and mechanism, overview
-
Spi
-
specific enzyme pro-peptide analogue inhibitor, purification from Streptococcus pyogenes, secondary structure and SpeB binding analysis, inhibition modeling and mechanism, overview
-
sulfite
-
-
tosyl-L-lysine chloromethyl ketone
-
irreversible inhibitor
tosyl-L-phenylalanine chloromethyl ketone
-
irreversible inhibitor
Tosyl-norleucine-Phe
-
competitive against benzyloxycarbonyl-norleucine-Phe
trans-epoxysuccinyl-L-leucylamido(4-guanidino)butane
-
-
trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane
Z-LVG-CHN2
-
irreversible inhibitor
Zn2+
-
Zinc inhibits the proteolytic activity of mature SpeB
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
dithiothreitol
-
reduction is essential for catalytic activity
Reducing agent
-
once the enzyme is activated by a reducing agent, the presence of the activator is no longer necessary for the catalytic activity
-
thioethanol
-
reduction is essential for catalytic activity, once the enzyme is activated by the reducing agent, the presence of the activator is no longer necessary for catalytic activity
thioglycolic acid
-
reduction is essential for catalytic activity
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.1178 - 0.2477
acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin
3.88
benzyloxycarbonyl-Glu-Phe
-
-
0.22 - 0.494
benzyloxycarbonyl-Lys-p-nitrophenyl ester
2.28
benzyloxycarbonyl-Lys-Phe
-
-
0.193
benzyloxycarbonyl-Val-Arg-4-methylcoumarinyl-7-amide hydrochloride
-
-
-
0.334
cinnamoyl-Gly-p-nitrophenyl ester
-
-
0.422
N-benzoyl-Gly-p-nitrophenyl ester
-
-
0.132
N-benzyloxycarbonyl-Ala-p-nitrophenyl ester
-
-
0.0236
N-benzyloxycarbonyl-Gly-p-nitrophenyl ester
-
-
0.588 - 0.667
Nalpha-benzyloxycarbonyl-Lys-p-nitrophenyl ester
5.05
Nalpha-benzyloxycarbonyl-Lys-Phe
-
-
-
2.64
Nalpha-benzyloxycarbonyl-Nepsilon-t-butyloxycarbonyl-Lys-Phe
-
-
0.063
p-nitrobenzyloxycarbonyl-Gly-p-nitrophenyl ester
-
-
0.438
t-butyloxycarbonyl-Gly-p-nitrophenyl ester
-
-
-
0.61
tert-butyloxycarbonyl-Ala-p-nitrophenyl ester
-
-
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.35 - 2.14
acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin
2.3
benzyloxycarbonyl-Glu-Phe
Streptococcus sp.
-
-
29.7 - 142
benzyloxycarbonyl-Lys-p-nitrophenyl ester
1.09
benzyloxycarbonyl-Lys-Phe
Streptococcus sp.
-
-
0.042
benzyloxycarbonyl-Val-Arg-4-methylcoumarinyl-7-amide hydrochloride
Streptococcus pyogenes
-
-
-
1.52
cinnamoyl-Gly-p-nitrophenyl ester
Streptococcus sp.
-
-
0.21
N-benzoyl-Gly-p-nitrophenyl ester
Streptococcus sp.
-
-
118
N-benzyloxycarbonyl-Ala-p-nitrophenyl ester
Streptococcus sp.
-
-
9.4
N-benzyloxycarbonyl-Gly-p-nitrophenyl ester
Streptococcus sp.
-
-
176 - 190
Nalpha-benzyloxycarbonyl-Lys-p-nitrophenyl ester
5.05
Nalpha-benzyloxycarbonyl-Lys-Phe
Streptococcus sp.
-
-
-
174
Nalpha-benzyloxycarbonyl-Lys-phenyl ester
Streptococcus sp.
-
at pH 7.25
8.6
Nalpha-benzyloxycarbonyl-Nepsilon-t-butyloxycarbonyl-Lys-Phe
Streptococcus sp.
-
-
1.15
p-nitrobenzyloxycarbonyl-Gly-p-nitrophenyl ester
Streptococcus sp.
-
-
0.147
t-butyloxycarbonyl-Gly-p-nitrophenyl ester
Streptococcus sp.
-
-
-
4.31
tert-butyloxycarbonyl-Ala-p-nitrophenyl ester
Streptococcus sp.
-
-
additional information
additional information
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.9 - 8.6
acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
-
inhibition kinetics with Spi and Spe B pro-peptide
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0279
Zn2+
Streptococcus pyogenes
-
pH and temperature not specified in the publication
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
341
-
protease activity substrate azocasein
378
-
protease activity substrate proSPE B C47S mutant
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
12000
-
pro-sequence, gel filtration
33590
-
determination of amino acid composition
35000
-
SDS-PAGE
42000
-
enzymatically inactive zymogen form of SpeB
160000
-
gel filtration
additional information
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
side-chain modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
analysis of a crystal structure model of the zymogen
-
crystal structures are determined of mature apo-SpeB at 1.06 A resolution from a crystal obtained in neutral pH conditions and high resolution crystal structures of SpeB in complex with the general proteinase inhibitor trans-epoxysuccinyl- L-leucylamido(4-guanidino)butane (E64) and in complex with the SpeB-specific N-terminal acetylated peptide aldehyde inhibitor acetyl-Ala-Gln-Ile-(S)-2,6-diaminohexanal
-
hanging drop vapour diffusion method, the crystal structure of the C47S mutant zymogen 40000 Da form expressed in Escherichia coli and its SeMet-substituted form, at 1.6 A resolution, reveals that the protein is a distant homologue of the papain superfamily
-
purified recombinant mutant C47S, X-ray diffraction crystal structure determination and analysis at 1.6 A resolution
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5 - 9.5
-
unstable below pH 4.5 and above pH 9.5
81451
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
0.1 N HCl, 2 h, denaturation of the zymogen
60
-
irreversible inactivation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
0.1 N HCl or 0.01 N NaOH, room temperature, 2 h, denaturation
-
0.1 N HCl, 2 h, denaturation of the zymogen
-
0.5% SDS, denaturation
-
8 M urea plus reducing agent, 20% of proteolytic activity is provided
-
EDTA, 0.001 M, stabilizes
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, stable for at least 2 years
-
stored under nitrogen, 0.01 mM EDTA, 3-5% loss of activity per week
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
HiTrap SP column chromatography, and G75 Sephadex gel filtration
-
native mature Spe B by ammonium sulfate fractionation, dialysis, and ion exchange chromatography, recombinant His-tagged Spe B propeptide and SpeB mutant C47S from Escherichia coli
-
Ni2+-chelating column chromatography
-
recombinant His-tagged wild-type and mutant C192S enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, the wild-type enzyme autoprocesses during purification to the mature 28 kDa protein
-
recombinant pro-sequence domain and refolded mature enzyme from Escherichia coli by affinity chromatography
-
recombinant wild-type and inactive mutant enzyme from Escherichia coli by affinity chromatography
-
the proteins are purified by Ni2+-chelating chromatography with a gradient of 20-200 mM imidazole. The proteins are concentrated by ultrafiltration using a 10 kDa cutoff membrane and then exchanged with phosphate-buffered saline
-
using Ni-NTA chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
C192S mutant expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli
-
expressed in Escherichia coli as a His-tagged fusion protein
-
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli BL21(DE3)pLyS cells
-
expression in Escherichia coli, the zymogen proSCP is secreted to the Escherichia coli periplasm followed by subsequent cleavage of the signal peptide
-
expression of 12 kDa pro-sequence domain of the enzyme in Escherichia coli, expression of mature enzyme in Escherichia coli in inclusion bodies
-
expression of His-tagged wild-type and mutant C192S enzymes in Escherichia coli strain BL21(DE3)
-
expression of mutant C47S
-
expression of wild-type and inactive mutant enzyme in Escherichia coli strain BL21(DE3) and in A-549 cells
-
gene speB, native expression pattern, expression of wild-type and mutant enzymes in Escherichia coli strain DH5alpha
-
genes speB and spi, nucleotide and amino acid sequence determination, analysis and comparison, co-expression of immature Spe B and Spi in Streptococcus pyogenes, Spi is similar to the pro-peptide of Spe B, overview, overexpression of His-tagged Spe B propeptide and SpeB mutant C47S in Escherichia coli
-
IdeS GST-fusion protein is expressed in Escherichia coli BL21
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D9N
-
protease activity (microgram/min/mg) substrate azocasein: 256, substrate proSPE B C47S mutant: 250, compared to wild-type 341 and 378, respectively
G136A
-
protease activity (microgram/min/mg) substrate azocasein: 22.3, substrate proSPE B C47S mutant: 8.1, compared to wild-type 341 and 378, respectively
G163S
-
protease activity (microgram/min/mg) substrate azocasein: 348, substrate proSPE B C47S mutant: 385, compared to wild-type 341 and 378, respectively
G163S/A172S
-
protease activity (microgram/min/mg) substrate azocasein: 286, substrate proSPE B C47S mutant: 236, compared to wild-type 341 and 378, respectively
G239D
-
protease activity (microgram/min/mg) substrate azocasein: 35.3, substrate proSPE B C47S mutant: 20, compared to wild-type 341 and 378, respectively
G308S
-
behaviour is similar to the wild type enzyme
G378A
-
Km(acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) and kcat (acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) decreased compared to wild-type
G380A
-
Km(acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) and kcat (acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) decreased compared to wild-type
G381A
-
Km(acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) and kcat (acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) decreased compared to wild-type
G382A
-
Km(acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) and kcat (acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) decreased compared to wild-type
G384A
-
Km(acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) and kcat (acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) decreased compared to wild-type
G384D
-
Km(acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) decreased compared to wild-type and kcat (acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) 50% decreased compared to wild-type
G385A
-
Km(acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) minimally increased compared to wild-type and kcat (acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) 3fold increased
T379A
-
Km(acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) and kcat (acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) slightly decreased compared to wild-type
T379V
-
Km(acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) and kcat (acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) slightly decreased compared to wild-type
V189A
-
protease activity (microgram/min/mg) substrate azocasein: 2.6, substrate proSPE B C47S mutant: 0.9, compared to wild-type 341 and 378, respectively
W212A
-
protease activity (microgram/min/mg) substrate azocasein: 1.2, substrate proSPE B C47S mutant: 0.9, compared to wild-type 341 and 378, respectively
W214A
-
protease activity (microgram/min/mg) substrate azocasein: 0.8, substrate proSPE B C47S mutant: 0.8, compared to wild-type 341 and 378, respectively
C192S
-
inactive mutant
-
G378A
-
Km(acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) and kcat (acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) decreased compared to wild-type
-
G380A
-
Km(acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) and kcat (acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) decreased compared to wild-type
-
G381A
-
Km(acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) and kcat (acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) decreased compared to wild-type
-
G382A
-
Km(acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) and kcat (acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) decreased compared to wild-type
-
G384D
-
Km(acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) decreased compared to wild-type and kcat (acetyl-Ala-Ile-Arg-7-amino-4-methylcoumarin) 50% decreased compared to wild-type
-
C47S
-
site-directed mutagenesis, inactive mutant
-
additional information
-
construction of an isogenic inactive mutant strain M6
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
90% renaturation provided the denatured zymogen is solubilized with urea or guanidinium chloride and given time to refold
-
refolding of denatured recombinant mature wild-type enzyme from inclusion bodies in 100 mM Na-acetate, pH 4.5, 8 M urea, 10 mM DTT, and 280 mM NaCl, by rapid dilution 1:20 into phosphate buffered saline, pH 7.4, with 10 mM DTT
-