Information on EC 3.4.22.47 - gingipain K

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The expected taxonomic range for this enzyme is: Porphyromonas gingivalis

EC NUMBER
COMMENTARY hide
3.4.22.47
-
RECOMMENDED NAME
GeneOntology No.
gingipain K
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
endopeptidase with strict specificity for lysyl bonds
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
159745-69-4
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain ATCC33277
-
-
Manually annotated by BRENDA team
strain H66
-
-
Manually annotated by BRENDA team
strain HG66
-
-
Manually annotated by BRENDA team
strain W83
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
-
Porphyromonas gingivalis Lys-gingipain (Kgp) facilitates heme acquisition by HmuY (heme-binding protein)
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
acetyl-lysine-4-nitroanilide + H2O
acetyl-lysine + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
acid-soluble human placental type I collagen + H2O
fragments of acid-soluble human placental type I collagen
show the reaction diagram
-
-
-
-
-
adrenocorticotropic hormone fragment 11-24 + H2O
fragments of adrenocorticotropic hormone fragment 11-24
show the reaction diagram
-
-
-
?
benzoxycarbonyl-L-lysine-4-nitroanilide + H2O
benzoxycarbonyl-L-lysine + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-L-Lys-p-nitroanilide + H2O
benzyloxycarbonyl-L-Lys + p-nitroaniline + H2O
show the reaction diagram
beta-endorphin + H2O
fragments of beta-endorphin
show the reaction diagram
-
-
-
?
bovine hemoglobin + H2O
fragments of bovine hemoglobin
show the reaction diagram
-
-
-
-
-
bovine serum albumin + H2O
fragments of bovine serum albumin
show the reaction diagram
-
-
-
-
-
bradykinin + H2O
fragments of bradykinin
show the reaction diagram
casein + H2O
fragments of casein
show the reaction diagram
-
-
-
-
-
CBZ-Phe-Arg-4-methyl-coumaryl-7-amide + H2O
?
show the reaction diagram
-
-
-
-
?
CD46 + H2O
?
show the reaction diagram
-
recombinant CD46 is degraded by Lys-gingipain in a dose-dependent manner
-
-
?
D-Val-Leu-Lys-p-nitroanilide + H2O
D-Val-Leu-Lys + p-nitroaniline
show the reaction diagram
D-Val-Phe-Lys-p-nitroanilide + H2O
D-Val-Phe-Lys + p-nitroaniline
show the reaction diagram
elafin + H2O
?
show the reaction diagram
all three gingipains have the ability to degrade elafin (endogenous inhibitor secreted by epithelial cells)
-
-
?
Fibrinogen + H2O
Fragments of fibrinogen
show the reaction diagram
focal adhesion kinase + H2O
?
show the reaction diagram
-
Kgp is responsible for degradation of integrin-related molecules
-
-
?
haptoglobin + H2O
fragments of haptoglobin
show the reaction diagram
-
human serum haptoglobin
-
?
Hemoglobin + H2O
?
show the reaction diagram
hemoglobin + H2O
fragments of hemoglobin
show the reaction diagram
-
human serum hemoglobin
-
?
hemopexin + H2O
fragments of hemopexin
show the reaction diagram
-
human serum hemopexin
-
?
human beta-defensin 3 + H2O
?
show the reaction diagram
-
-
-
-
?
human IgA + H2O
cleaved human IgA
show the reaction diagram
-
-
-
-
-
human IgG + H2O
fragments of human IgG
show the reaction diagram
-
-
-
-
-
IgG1 + H2O
?
show the reaction diagram
-
The heavy chain of IgG1 is cleaved at a single site within the hinge region, generating Fab and Fc fragments
-
-
?
IgG3 + H2O
?
show the reaction diagram
-
IgG3 is cleaved within the heavy chain and at several sites around the CH2 region
-
-
?
IL-6 + H2O
fragments of IL-6
show the reaction diagram
-
-
-
?
interleukin 8 + H2O
fragments of interleukin 8
show the reaction diagram
interleukin-1beta + H2O
?
show the reaction diagram
interleukin-6 + H2O
?
show the reaction diagram
interleukin-8 + H2O
?
show the reaction diagram
melittin + H2O
fragments of melittin
show the reaction diagram
-
-
-
?
MeoSuc-Ala-Ala-Pro-Val-4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
?
Met-Lys-bradykinin + H2O
fragments of Met-Lys-bradykinin
show the reaction diagram
-
-
-
?
methemoglobin + H2O
?
show the reaction diagram
monocyte chemotactic protein 1 + H2O
fragments of monocyte chemotactic protein 1
show the reaction diagram
N-(p-tosyl)-Gly-Pro-Lys 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
N-(p-tosyl)-Gly-Pro-Lys-4-nitroanilide + H2O
N-(p-tosyl)-Gly-Pro-Lys + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
N-4-tosyl-Gly-Pro-Lys-4-nitroanilide + H2O
N-4-tosyl-Gly-Pro-Lys + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
N-alpha-acetyl-L-lysine-4-nitroanilide + H2O
N-alpha-acetyl-L-lysine + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
N-alpha-acetyl-L-lysine-p-nitroanilide + H2O
N-alpha-acetyl-L-lysine + 4-nitroaniline
show the reaction diagram
-
-
-
-
r
N-alpha-benzoyl-DL-lysine 4-nitroanilide + H2O
N-alpha-benzoyl-DL-lysine + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
N-alpha-benzyloxycarbonyl-L-lysine-4-nitroanilide + H2O
N-alpha-benzyloxycarbonyl-L-lysine + 4-nitroaniline
show the reaction diagram
N-p-tosyl-Gly-Pro-Lys-4-nitroanilide + H2O
N-p-tosyl-Gly-Pro-Lys + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
N-p-tosyl-Gly-Pro-Lys-p-nitroanilide + H2O
N-p-tosyl-Gly-Pro-Lys + p-nitroaniline
show the reaction diagram
Nalpha-acetyl-L-lysine 4-nitroanilide + H2O
Nalpha-acetyl-L-lysine + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
neurotensin + H2O
Glu-Leu-Tyr-Glu-Asn-Lys + Arg-Arg-Pro-Tyr-Ile-Leu
show the reaction diagram
osteoprotegerin + H2O
?
show the reaction diagram
oxyhaemoglobin + H2O
?
show the reaction diagram
-
-
-
-
?
paxillin + H2O
?
show the reaction diagram
-
Kgp is responsible for degradation of integrin-related molecules
-
-
?
protease-activated receptor-2 + H2O
?
show the reaction diagram
-
specific substrate for Lys-gingipain
-
-
?
t-butyl-oxycarbonyl-L-Glu-L-Lys-L-Lys-4-methyl-7-coumarylamide + H2O
t-butyl-oxycarbonyl-L-Glu-L-Lys-L-Lys + 7-amino-4-methylcoumarin
show the reaction diagram
-
41% of activity with t-butyl-oxycarbonyl-L-Val-L-Leu-L-Lys-4-methyl-7-coumarylamide
-
?
t-butyl-oxycarbonyl-L-Val-L-Leu-L-Lys-4-methyl-7-coumarylamide + H2O
t-butyl-oxycarbonyl-L-Val-L-Leu-L-Lys + 7-amino-4-methylcoumarin
show the reaction diagram
-
gingipain K has a stron preference for substrates containing Lys in the p1 site
-
?
t-butyloxycarbonyl-Val-Leu-Lys-Phe-Arg-4-methyl-7-coumarylamide
t-butyl-oxycarbonyl-Val-Leu-Lys + Phe-Arg-4-methyl-7-coumarylaminde
show the reaction diagram
-
-
-
-
?
thrombomodulin + H2O
?
show the reaction diagram
-
Lys-gingipain and Arg-gingipain cleave thrombomodulin in vitro
-
-
?
TNFalpha + H2O
?
show the reaction diagram
transferrin + H2O
fragments of transferrin
show the reaction diagram
-
human serum transferrin
-
?
Val-Leu-Lys-4-nitroanilide + h2O
Val-Leu-Lys + 4-nitroaniline
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
bradykinin + H2O
fragments of bradykinin
show the reaction diagram
-
together with gingipain R, gingipain K induces vascular permeability enhancement in human plasma by cleaving bradykinin from high molecular weight kininogen
-
-
-
elafin + H2O
?
show the reaction diagram
B2RLK2
all three gingipains have the ability to degrade elafin (endogenous inhibitor secreted by epithelial cells)
-
-
?
Fibrinogen + H2O
Fragments of fibrinogen
show the reaction diagram
-
most prominent target among plasma proteins
-
-
-
focal adhesion kinase + H2O
?
show the reaction diagram
-
Kgp is responsible for degradation of integrin-related molecules
-
-
?
Hemoglobin + H2O
?
show the reaction diagram
-
degradation, the enzyme forms a complex with the outer membrane receptor HmuR required for binding and utilization of hemoglobin and hemin, overview
-
-
?
interleukin 8 + H2O
fragments of interleukin 8
show the reaction diagram
interleukin-1beta + H2O
?
show the reaction diagram
-
biological inactivation and degradation, low activity, cytokine degradation is mainly the result of Lys-gingipain
-
-
?
interleukin-6 + H2O
?
show the reaction diagram
-
biological inactivation and degradation
-
-
?
interleukin-8 + H2O
?
show the reaction diagram
-
biological inactivation and degradation
-
-
?
methemoglobin + H2O
?
show the reaction diagram
-
degradation, ligation of methemoglobin with N3-, which effectively blocks heme dissociation from the protein, prevents hemoglobin breakdown
-
-
?
monocyte chemotactic protein 1 + H2O
fragments of monocyte chemotactic protein 1
show the reaction diagram
-
degradation of human monocyte chemotactic protein 1 in Porphyromonas gingivalis-infected human umbilical vein endothelial cells
-
-
-
osteoprotegerin + H2O
?
show the reaction diagram
paxillin + H2O
?
show the reaction diagram
-
Kgp is responsible for degradation of integrin-related molecules
-
-
?
protease-activated receptor-2 + H2O
?
show the reaction diagram
-
specific substrate for Lys-gingipain
-
-
?
TNFalpha + H2O
?
show the reaction diagram
-
degradation, leading to inhibition of biological functions of TNFalpha, overview
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Zn2+
-
enhances the inhibitory effect of chlorhexidine
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-(3-phenylpropionyl)piperidine-3-(R,S)-carboxylic acid-[4-amino-1(S)-(benzothiazole-2-carbonyl)butyl] amide
-
reversible inhibition
benzamidine derivatives
-
-
benzyloxycarbonyl-L-phenylalanyl-L-lysyl-acyloxyketone
-
-
benzyloxycarbonyl-L-phenylalanyl-L-lysylacyloxyketone
-
the inhibitor completely abolishes osteoclastogenesis induced by Kgp
benzyloxycarbonyl-Phe-Lys-chloromethylketone
-
i.e. z-FKck, specific for Kgp
carbobenzoxy-Glu(NHN(CH3)Ph)-Lys-CO-NHCH2Ph
-
-
carbobenzoxy-Lys-Arg-CO-Lys-N-(CH3)2
-
-
cathepsin B inhibitor
-
-
-
Chlorhexidine
-
synergistic effect of Zn2+ in a 1:1 ratio of chlorhexidine and Zn2+
Chloromethyl ketones
-
development of diverse inhibitor derivatives: structure-based design, chemistry, and activity, specificity for the Sn binding pocket of the enzyme, overview
Chloromethylketones
-
-
CuSO4
-
1 mM, 79% inhibition
FeCl3
-
1 mM, 42% inhibition
Gly-Gly
-
200 mM, 50% inhibition
iodoacetamide
iodoacetic acid
-
1 mM, 76% inhibition
kappa-casein
-
inhibits proteolytic activity associated with Porphyromonas gingivalis whole cells, purified RgpA-Kgp proteinase-adhesin complexes, and purified RgpB proteinase. The peptide kappa-casein(109-137) exhibits synergism with Zn(II) against both Arg- and Lys-specific proteinases. Active region for inhibition is identified as kappa-casein (117-137). Kappa-casein inhibits in an uncompetitive manner
-
KYT-36
lactoferrin
-
inhibits both the Arg- and Lys-specific proteinase activities of Porphyromonas gingivalis whole cells by approximately 40% at 1 mg/ml and over 70% at 10 mg/ml. Lactoferrin inhibits both the Arg-specific and Lys-specific activities of purified Porphyromonas gingivalis 248 RgpA/Kgp proteinase-adhesin complexes by 96% at a concentration of 5 mg/mL
-
Leupeptin
lysine
-
slight inhibition of coaggregation of Porphyromonas gingivalis with other oral bacteria by L-lysine and more slightly by D-lysine
MnSO4
-
1 mM, 50% inhibition
N-alpha-p-tosyl-L-lysine chloromethyl ketone
-
0.1 mM, complete inhibition
N-ethylmaleimide
Nalpha-benzoyl-Phe-Lys-chloromethyl ketone
-
a specific inhibitor of lysine gingipain
p-hydroxymercuribenzoate
-
0.2 mM, 68% inhibition
Phe-Pro-Arg-chloromethyl ketone
-
0.1 mM, 95% inhibition
porcine pancreatic secretory trypsin inhibitor
-
i.e. PSTI porcine, a Kazal-type serine proteinase inhibitor purified from pancreas, porcine pancreatic secretory trypsin inhibitor having an essential Lys residue at the P1 position of the reactive site, and containing Tyr and Asn residues the P2' and P3' sites, specifically inhibits the activity of the Lys-specific gingipain K, whereas bovine inhibitor, possessing a Arg residue at the P1 position, exhibits activity only against the Arg-specific cysteine proteinase gingipain K, EC 3.4.22.37. The association equilibrium constant is 0.51 mM
-
Pro-Phe-Arg-chloromethylketone
-
-
rice grain extract
-
a rice protein fraction is shown to have Rgp inhibitory activities. Comprehensive affinity chromatography and MS analyses results in the identification of 4 proteins a 26 kDa globulin, a plant lipid transfer/trypsin-alpha amylase inhibitor, the RA17 seed allergen, and an alpha amylase/trypsin inhibitor proteins accounting for 90% of the inhibitory activity. Inhibitory activity against Rgp is 20fold higher than that against Kgp
-
tetracycline
-
-
tosyl-L-lysine chloromethyl ketone
tosyl-L-phenylalanine chloromethyl ketone
-
1 mM, 99% inhibition
tosyl-Lys-chloromethylketone
-
-
Z-Phe-Lys-2,4,6-trimethyl-benzoyloxymethyl-ketone
-
specific inhibition of Kgp
Z-Phe-Lys-acyloxymethylketone
-
oxyhemoglobin-Kgp interactions resulting in formation of a hemoglobin hemichrome are inhibited by specific inhibitor Z-Phe-Lys-acyloxymethylketone
zFKck
-
human gingvial epithelial cells treated with Kgp-specific inhibitor leupeptin and challenged with Porphyromonas gingivalis cells do not undergo apoptosis
ZnCl2
-
1 mM, 50% inhibition
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
cysteine
dithiothreitol
EDTA
-
2 mM, 79% increase in activity
EGTA
-
2 mM, 79% increase in activity
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.18
benzyloxycarbonyl-L-Lys-p-nitroanilide
-
pH 8.5, 37°C
0.2
D-Val-Leu-Lys-p-nitroanilide
-
pH 8.5, 37°C
0.126
D-Val-Phe-Lys-p-nitroanilide
-
pH 8.5, 37°C
0.0029
hemoglobin
-
pH 7.6, 37°C, human serum hemoglobin
-
0.05
N-p-tosyl-Gly-Pro-Lys-p-nitroanilide
-
pH 8.5, 37°C
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
9.4
hemoglobin
Porphyromonas gingivalis
-
pH 7.6, 37°C, human serum hemoglobin
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0000009
1-(3-phenylpropionyl)piperidine-3-(R,S)-carboxylic acid-[4-amino-1(S)-(benzothiazole-2-carbonyl)butyl]amide
-
-
additional information
additional information
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.26
-
substrate N-p-tosyl-Gly-Pro-Lys-p-nitroanilide
0.356
-
release of 7-amino-4-methylcoumarin
additional information
-
enzyme activity and hemoglobin/hemin binding in wild-type strain A7436 and in kgp-deficient mutant strains WS1, WS10, and WS15, overview
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.6 - 8.3
-
assay at
8.5
-
with protein substrates like azocasein
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 9.5
-
hydrolysis of t-butyl-oxycarbonyl-L-Val-L-Leu-L-Lys-4-methyl-7-coumarylamide
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
Porphyromonas gingivalis secretes outer membrane vesicles that contain major virulence factors, including Arg-gingipain and Lys-gingipain
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50000
-
SDS-PAGE
52000
-
gel filtration
105000
-
gel filtration without boiling
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
-
1 * 51000, SDS-PAGE
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
the crystal structure of a domain within the haemagglutinin region of Kgp is reported here. The K2 domain structure is a jellyroll fold with two anti-parallel beta-sheets. This fold topology is shared with adhesive domains from functionally diverse receptors such as MAM domains, ephrin receptor ligand binding domains and a number of carbohydrate binding modules
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.5
-
40% loss of activity after 5 min
638848
5 - 9
-
stable for several hours in the absence of cysteine, rapid loss of activity below pH 8.0 in the presence of cysteine
638843
5.5 - 10.5
-
0°C, no loss of activity
638848
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
no loss of activity
37
-
no loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
Kgp shows a loss of only 1% and 8% of initial activity after 7 and 24 h, respectively
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
acetone precipitate, Sephadex G-150, arginine-Sepharose
-
ammonium sulfate, immunoaffinity column, Mono S
-
ammonium sulfate, Mono Q, chromatofocusing on MonoP
-
Lys-gingipain is purified from the cell envelope extract of Porphyromonas gingivalis KDP112 by affinity chromatography on arginine Sepharose 4B
-
native extracellular enzyme from culture medium
-
native Kgp from culture supernatant
-
native Lys-gingipain from cell culture supernatant, re-activation of the proteases by reducing the active-centre thiol group partially oxidized during the purification process
-
purified by using gel-filtration and arginine-Sepharose chromatography
-
using gel-filtration
-
using Ni-NTA chromatography and gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Sf9 insect cells
expression of wild-type and mutant enzymes in murine ST2 osteoblastic/stromal cells
-
HbR domain is recombinantly expressed in Escherichia coli
-
the gene fragment encoding the K2 domain (Ala1157-Gly1334) of Kgp from Porphyromonas gingivalis W83 is cloned into a modified vector pET-32a. The constructed plasmid is transformed into Escherichia coli BL21(DE3) competent cells for protein expression
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C476A
-
high Kgp activity and black pigmentation
C476A/C477A
-
no Kgp activity nor black pigmentation
C477A
-
no Kgp activity nor black pigmentation
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
molecular biology