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Information on EC 3.4.21.B4 - granzyme K and Organism(s) Homo sapiens and UniProt Accession P49863
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3.4.21.B4 granzyme KSpecify your search results
The taxonomic range for the selected organisms is: Homo sapiens
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Synonyms
granzyme k, fragmentin, granzyme 3, rnk-tryp-2, gzm k, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
GzmK
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of peptide bond
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-
CAS REGISTRY NUMBER
COMMENTARY
196622-94-3
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
benzyloxycarbonyl-Arg p-nitrobenzyl ester + H2O
benzyloxycarbonyl-Arg + p-nitrobenzyl alcohol
-
-
-
?
benzyloxycarbonyl-L-Lys-thiobenzyl ester + H2O
benzyloxycarbonyl-L-Lys + thiobenzyl alcohol
-
-
-
?
beta-tubulin + H2O
?
cleaves beta-tubulin after Arg62 (YVPR-/-AV) and Arg282 (QQYR-/-AL)
-
-
?
Cbz-L-Arg-4-nitroanilide + H2O
Cbz-L-Arg + 4-nitroaniline
little hydrolyzed by Gr3
-
-
?
Cbz-L-Arg-7-amido-4-methylcoumarin + H2O
Cbz-L-Arg + 7-amino-4-methylcoumarin
little hydrolyzed by Gr3
-
-
?
Cbz-L-Lys-4-nitroanilide + H2O
Cbz-L-Lys + 4-nitroaniline
little hydrolyzed by Gr3
-
-
?
fluorescence resonance energy transfer substrate-25Arg peptide + H2O
?
most effectively hydrolyzes the substrate fluorescence resonance energy transfer substrate-25Arg peptide in the FRETS-25Xaa series tested
-
-
?
fluorescence resonance energy transfer substrate-25Lys peptide + H2O
?
far less effectively cleaved than fluorescence resonance energy transfer substrate-25Arg peptide
-
-
?
heterogeneous nuclear ribonucleoprotein K + H2O
?
granzyme K and granzyme A cleave with different kinetics at distinct sites
-
-
?
Z-Leu-Arg-Gly-Gly-7-amido-4-methylcoumarin + H2O
Z-Leu-Arg + Gly-Gly-7-amino-4-methylcoumarin
-
-
-
?
benzyloxycarbonyl-L-Arg-thiobenzyl ester + H2O
benzyloxycarbonyl-L-Arg + thiobenzyl alcohol
-
-
-
-
?
benzyloxycarbonyl-Lys-thiobenzyl ester + H2O
benzyloxycarbonyl-Lys + thiobenzyl alcohol
-
-
-
-
?
Bid protein + H2O
tBid protein + ?
-
GzmK directly processes Bid to produce its active form tBid
-
-
?
Cys-Gly-Tyr-Gly-Pro-Lys-Lys-Lys-Arg-Lys-Val-Gly-Gly + H2O
Cys-Gly-Tyr-Gly-Pro-Lys + Lys-Lys-Arg + Lys-Val-Gly-Gly
-
-
-
-
?
redox factor-1/apurinic apyrimidinic endonuclease Ape1 + H2O
?
-
Ape1 cleavage by Granzyme K facilitates intracellular reactive oxygen species accumulation and enhances granzyme K-induced cell death
-
-
?
additional information
?
-
most efficiently hydrolyzes the carboxylic side of Phe-Tyr-Arg (P3-P2-P1) sequence
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-
?
additional information
?
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-
most efficiently hydrolyzes the carboxylic side of Phe-Tyr-Arg (P3-P2-P1) sequence
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-
?
additional information
?
-
granzyme K prefers Arg over Lys at P1 position. Granzyme K and granzyme A display highly restricted substrate specificities that overlapp only partially. Whereas granzyme K and granzyme A cleave SET with similar efficiencies likely at the same sites, both granzymes cleave the pre-mRNA-binding protein heterogeneous ribonuclear protein K with different kinetics at distinct sites. Granzyme K is markedly more efficient in cleaving heterogeneous ribonuclear protein K than granzyme A. Granzyme K, but not granzyme A, cleaves the microtubule network protein beta-tubulin after two distinct Arg residues. Neither GrK cleavage sites in beta-tubulin nor a peptide based proteomic screen reveal a clear granzyme K consensus sequence around the P1 residue, suggesting that GrK specificity depends on electrostatic interactions between exosites of the substrate and the enzyme
-
-
?
additional information
?
-
substrate identification analysing substrate repertoire and specificity by N-terminal COFRADIC assisted N-terminomics on the homologous enzyme. Comparisons of specificity profiles of human and mouse enzymes revealing only subtle differences and pointed to substrate occupancies in the P1, P1', and P2' position as the main determinants for substrate recognition, overview
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-
?
additional information
?
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-
substrate identification analysing substrate repertoire and specificity by N-terminal COFRADIC assisted N-terminomics on the homologous enzyme. Comparisons of specificity profiles of human and mouse enzymes revealing only subtle differences and pointed to substrate occupancies in the P1, P1', and P2' position as the main determinants for substrate recognition, overview
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-
?
additional information
?
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-
cleaves synthetic thiobenzyl ester substrates after Lys and Arg, no substrates for the mature enzyme: Boc-Ala-Ala-Met-thiobenzyl ester and Boc-Ala-Ala-Asp-thiobenzyl ester
-
?
additional information
?
-
-
for single-residue thioester substrates the enzyme shows a 2fold preference for a Lys versus Arg residue at P1. With oligopeptide substrates the enzyme displays peptidolytic activity C-terminal to both Lys and Arg residues with comparable rates of hydrolysis
-
?
additional information
?
-
-
can trigger DNA fragmentation and is involved in apoptosis
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
redox factor-1/apurinic apyrimidinic endonuclease Ape1 + H2O
?
-
Ape1 cleavage by Granzyme K facilitates intracellular reactive oxygen species accumulation and enhances granzyme K-induced cell death
-
-
?
additional information
?
-
-
can trigger DNA fragmentation and is involved in apoptosis
-
?
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
inter-alpha-inhibitor protein
physiological inhibitor, significantly inhibits catalytic activity in vitro
-
bikunin
-
subunit of inter-alpha-trypsin inhibitor complex, physiologic inhibitor
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.0529
cell extract, using benzyloxycarbonyl-Lys-thiobenzyl ester as a substrate, at 25°C
37.1
after 1202.9fold purification, using benzyloxycarbonyl-Arg p-nitrobenzyl ester as a substrate, at 25°C
63.7
after 1202.9fold purification, using benzyloxycarbonyl-Lys-thiobenzyl ester as a substrate, at 25°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
10.2
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
from patients with allergic asthma (before and after segmental allergen challenge), and in patients with mild chronic obstructive pulmonary disease (COPD), pneumonia and in healthy controls. Expression of granzyme K is upregulated in acute airway inflammation, both in infectious and non-infectious diseases
granzyme K is expressed and secreted by cultured classically activated macrophages
granzyme serum levels are elevated in patients with autoimmune diseases and infections, including sepsis
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
physiological function
physiological function
granzymes are serine proteases released by cytotoxic lymphocytes to induce apoptosis in virus-infected cells and tumor cells. Granzymes also play a role in controlling inflammation. Granzyme K binds to Gram-negative bacteria, e.g. Escherichia coli BL21, Pseudomonas aeruginosa, and Neisseria meningitides, and their cell-wall component lipopolysaccharide. The enzyme synergistically enhances lipopolysaccharide-induced cytokine release in vitro from primary human monocytes and in vivo in a mouse model of lipopolysaccharide challenge. The synergistic effect of the enzyme and lipopolysacchride on monocytes is dependent on cluster of differentiation 14, but the extracellular effects are independent of the enzyme catalytic activity. The enzyme disaggregates lipopolysaccharides from micelles and augments LPS-CD14 complex formation, thereby likely boosting monocyte activation by lipopolysaccharides. The extracellular enzyme is a direct modulator of LPS-TLR4 signaling during the antimicrobial innate immune response, detailed overview
physiological function
the enzyme has a role in cytotoxic and inflammatory processes
physiological function
extracellular granzyme K proteolytically activates protease-activated receptor-1 leading to increased interleukin 6 and monocyte chemotactic protein 1 production in endothelial cells. Granzyme K also increases tumour necrosis factor alpha-induced inflammatory adhesion molecule expression
physiological function
in burn tissue, granzyme K expression is elevated compared with normal skin, with expression predominantly found in macrophages. Granzyme K is expressed and secreted by cultured human classically activated macrophages
physiological function
-
granzyme K induces pro-inflammatory cytokine release (interleukin-6, interleukin-8 and monocyte chemotactic protein-1) through the activation of protease-activated receptor-1, induces activation of both the ERK1/2 and p38 MAP kinase signaling pathways, and significantly increases fibroblast proliferation
physiological function
-
valosin-containing protein cleavage by granzyme K accelerates an endoplasmic reticulum stress leading to caspase-independent cytotoxicity of target tumor cells
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). GRAK_HUMAN
264
0
28882
Swiss-Prot
other Location (Reliability: 2)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25000
28000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
enzyme quickly loses catalytic activity when diluted and stored in solution, Tween-20 or bovine serum albumin prevent loss of activity
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
DE52 anion-exchange column chromatography, Q Sepharose column chromatography, hydroxylapatite column chromatography, and Sephadex G-75 gel filtration
recombinant wild-type and mutant enzymes by cation-exchange chromatography and affinity chromatography on Prot A/G beads, followed by dialysis
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Babe, L.M.; Yoast, S.; Dreyer, M.; Schmidt, B.F.
Heterologous expression of human granzyme K in Bacillus subtilis and characterization of its hydrolytic activity in vitro
Biotechnol. Appl. Biochem.
27
117-124
1998
Homo sapiens
Wilharm, E.; Tschopp, J.; Jenne, D.E.
Biological activities of granzyme K are conserved in the mouse and account for residual Z-Lys-SBzl activity in granzyme A-deficient mice
FEBS Lett.
459
139-142
1999
Homo sapiens, Mus musculus
Wilharm, E.; Parry, M.A.; Friebel, R.; Tschesche, H.; Matschiner, G.; Sommerhoff, C.P.; Jenne, D.E.
Generation of catalytically active granzyme K from Escherichia coli inclusion bodies and identification of efficient granzyme K inhibitors in human plasma
J. Biol. Chem.
274
27331-27337
1999
Homo sapiens
Hink-Schauer, C.; Estebanez-Perpina, E.; Wilharm, E.; Fuentes-Prior, P.; Klinkert, W.; Bode, W.; Jenne, D.E.
The 2.2-A crystal structure of human pro-granzyme K reveals a rigid zymogen with unusual features
J. Biol. Chem.
277
50923-50933
2002
Homo sapiens
Hameed, A.; Lowrey, D.M.; Lichtenheld, M.; Podack, E.R.
Characterization of three serine esterases isolated from human IL-2 activated killer cells
J. Immunol.
141
3142-3147
1988
Homo sapiens
Jackson, D.S.; Fraser, S.A.; Ni, L.M.; Kam, C.M.; Winkler, U.; Johnson, D.A.; Froelich, C.J.; Hudig, D.; Powers, J.C.
Synthesis and evaluation of diphenyl phosphonate esters as inhibitors of the trypsin-like granzymes A and K and mast cell tryptase
J. Med. Chem.
41
2289-2301
1998
Homo sapiens
Bratke, K.; Kuepper, M.; Bade, B.; Virchow, J.C.; Luttmann, W.
Differential expression of human granzymes A, B, and K in natural killer cells and during CD8+ T cell differentiation in peripheral blood
Eur. J. Immunol.
35
2608-2616
2005
Homo sapiens
Bade, B.; Lohrmann, J.; ten Brinke, A.; Wolbink, A.M.; Wolbink, G.J.; ten Berge, I.J.; Virchow, J.C.; Luttmann, W.; Hack, C.E.
Detection of soluble human granzyme K in vitro and in vivo
Eur. J. Immunol.
35
2940-2948
2005
Homo sapiens
Babe, L.M.; Schmidt, B.F.
Granzyme K
Handbook of Proteolytic Enzymes (Barrett, A. J. , Rawlings, N. D. , Woessner; j. F. , eds. )Academic Press
2
1552-1554
2004
Homo sapiens, Rattus norvegicus
-
Hirata, Y.; Inagaki, H.; Shimizu, T.; Li, Q.; Nagahara, N.; Minami, M.; Kawada, T.
Expression of enzymatically active human granzyme 3 in Escherichia coli for analysis of its substrate specificity
Arch. Biochem. Biophys.
446
35-43
2006
Homo sapiens (P49863), Homo sapiens
Zhao, T.; Zhang, H.; Guo, Y.; Zhang, Q.; Hua, G.; Lu, H.; Hou, Q.; Liu, H.; Fan, Z.
Granzyme K cleaves the nucleosome assembly protein SET to induce single-stranded DNA nicks of target cells
Cell Death Differ.
14
489-499
2007
Homo sapiens
Zhao, T.; Zhang, H.; Guo, Y.; Fan, Z.
Granzyme K directly processes Bid to release cytochrome c and endonuclease G leading to mitochondria-dependent cell death
J. Biol. Chem.
282
12104-12111
2007
Homo sapiens
Guo, Y.; Chen, J.; Zhao, T.; Fan, Z.
Granzyme K degrades the redox/DNA repair enzyme Ape1 to trigger oxidative stress of target cells leading to cytotoxicity
Mol. Immunol.
45
2225-2235
2008
Homo sapiens
Bovenschen, N.; Quadir, R.; van den Berg, A.L.; Brenkman, A.B.; Vandenberghe, I.; Devreese, B.; Joore, J.; Kummer, J.A.
Granzyme K displays highly restricted substrate specificity that only partially overlaps with granzyme A
J. Biol. Chem.
284
3504-3512
2009
Homo sapiens (P49863)
Bratke, K.; Klug, A.; Julius, P.; Kuepper, M.; Lommatzsch, M.; Sparmann, G.; Luttmann, W.; Virchow, J.C.
Granzyme K: a novel mediator in acute airway inflammation
Thorax
63
1006-1011
2008
Homo sapiens (P49863), Homo sapiens
Guo, Y.; Chen, J.; Shi, L.; Fan, Z.
Valosin-containing protein cleavage by granzyme K accelerates an endoplasmic reticulum stress leading to caspase-independent cytotoxicity of target tumor cells
J. Immunol.
185
5348-5359
2010
Homo sapiens
Cooper, D.M.; Pechkovsky, D.V.; Hackett, T.L.; Knight, D.A.; Granville, D.J.
Granzyme K activates protease-activated receptor-1
PLoS ONE
6
e21484
2011
Homo sapiens
Plasman, K.; Demol, H.; Bird, P.I.; Gevaert, K.; Van Damme, P.
Substrate specificities of the granzyme tryptases A and K
J. Proteome Res.
13
6067-6077
2014
Homo sapiens (P49863), Homo sapiens, Mus musculus
Wensink, A.C.; Kemp, V.; Fermie, J.; Garcia Laorden, M.I.; van der Poll, T.; Hack, C.E.; Bovenschen, N.
Granzyme K synergistically potentiates LPS-induced cytokine responses in human monocytes
Proc. Natl. Acad. Sci. USA
111
5974-5979
2014
Homo sapiens (P49863), Homo sapiens
Sharma, M.; Merkulova, Y.; Raithatha, S.; Parkinson, L.G.; Shen, Y.; Cooper, D.; Granville, D.J.
Extracellular granzyme K mediates endothelial activation through the cleavage of protease-activated receptor-1
FEBS J.
283
1734-1747
2016
Homo sapiens (P49863), Homo sapiens
Turner, C.T.; Zeglinski, M.R.; Richardson, K.C.; Zhao, H.; Shen, Y.; Papp, A.; Bird, P.I.; Granville, D.J.
Granzyme K expressed by classically activated macrophages contributes to inflammation and impaired remodeling
J. Invest. Dermatol.
139
930-939
2019
Homo sapiens (P49863), Homo sapiens, Mus musculus (O35205)
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