Information on EC 3.4.21.B2 - granzyme M

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.4.21.B2
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
granzyme M
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
endopeptidase activity. Cleaves substrates containing a methionine side chain at P1
show the reaction diagram
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-
-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
cleavage of C-N-linkage
-
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hydrolysis of peptide bond
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CAS REGISTRY NUMBER
COMMENTARY hide
159745-66-1
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
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a lack of GrzM results in reduced serum IL-1alpha, IL-1beta, TNF, and IFN-gamma levels and significantly reduces susceptibility to lipopolysaccharide-induced lethal endotoxicosis
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-GRLL-7-amido-4-methylcoumarin + H2O
acetyl-GRLL + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
acetyl-KEPL-7-amido-4-methylcoumarin + H2O
acetyl-KEPL + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
acetyl-KEPM-7-amido-4-methylcoumarin + H2O
acetyl-KEPM + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
acetyl-KVAM-7-amido-4-methylcoumarin + H2O
acetyl-KVAM + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
acetyl-KVPL-7-amido-4-methylcoumarin + H2O
acetyl-KVPL + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
acetyl-KVPM-7-amido-4-methylcoumarin + H2O
acetyl-KVPM + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
acetyl-KYAL-7-amido-4-methylcoumarin + H2O
acetyl-KYAL + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
acetyl-KYPL-7-amido-4-methylcoumarin + H2O
acetyl-KYPL + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
acetyl-KYPM-7-amido-4-methylcoumarin + H2O
acetyl-KYPM + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
acetyl-MEPL-7-amido-4-methylcoumarin + H2O
acetyl-MEPL + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
acetyl-RYPL-7-amido-4-methylcoumarin + H2O
acetyl-RYPL + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
acetyl-VPL-7-amido-4-methylcoumarin + H2O
acetyl-VPL + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
alpha-Tubulin + H2O
?
show the reaction diagram
ezrin + H2O
?
show the reaction diagram
-
granzyme M directly and efficiently cleaves the actin-plasma membrane linker ezrin
-
-
?
heterogeneous nuclear ribonucleoprotein K + H2O
?
show the reaction diagram
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-
granzyme M most efficiently cleaves heterogeneous nuclear ribonucleoprotein K in the presence of RNA at multiple sites, thereby likely destroying heterogeneous nuclear ribonucleoprotein K function
-
?
human Fas-associated protein with Death Domain + H2O
?
show the reaction diagram
inhibitor of caspase-activated DNase + H2O
?
show the reaction diagram
mouse Fas-associated protein with Death Domain mutant V193M/L194S + H2O
?
show the reaction diagram
N-benzyloxycarbonyl-(N-(beta-t-butylseryl))-prolyl-methionine-thiobenzylester + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-(N-(epsilon-t-butyloxycarbonyllysyl))-prolyl-methionine-thiobenzylester + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-(N-(gamma-t-butylaspartyl))-prolyl-methionine-thiobenzylester + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-alanyl-alanyl-methionine-thiobenzylester + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-alanyl-alanyl-prolyl-methionine-thiobenzylester + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-alanyl-aspartyl-methionine-thiobenzylester + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-alanyl-lysyl-methionine-thiobenzylester + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-alanyl-prolyl-methionine-thiobenzylester + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-alanyl-seryl-methionine-thiobenzylester + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-aspartyl-prolyl-methionine-thiobenzylester + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-L-Phe-L-Pro-L-Met-thiobenzyl ester + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-lysyl-prolyl-methionine-thiobenzylester + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-prolyl-prolyl-methionine-thiobenzylester + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-seryl-prolyl-methionine-thiobenzylester + H2O
?
show the reaction diagram
-
-
-
-
?
N-tert-butoxycarbonyl-Ala-Ala-Met thiobenzyl ester + H2O
N-tert-butoxycarbonyl-Ala-Ala-Met + phenylmethanethiol
show the reaction diagram
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activity only occurs with deletion mutants, but not with wild-type enzyme
-
?
N-tert-butoxycarbonyl-Ala-Ala-Met-thiobenzyl ester + H2O
N-tert-butoxycarbonyl-Ala-Ala-Met + phenylmethanethiol
show the reaction diagram
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cleaves thiobenzylester substrates specifically after methionine, norleucine or leucine residues in the primary substrate site P1
-
?
nucleophosmin + H2O
?
show the reaction diagram
poly(ADP-ribose) polymerase + H2O
?
show the reaction diagram
Protein + H2O
?
show the reaction diagram
proteinase inhibitor 9 + H2O
?
show the reaction diagram
-
effective hydrolysis and inactivation, bypassing proteinase inhibitor 9 inhibition of granzyme B
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-
?
succinyl-Ala-Ala-Pro-Leu-4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
survivin + H2O
?
show the reaction diagram
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the inhibitor of the apoptosis gene family member survivin is a physiological substrate for GzmM. GzmM hydrolyzes survivin at Leu-138 to remove the last four C-terminal residues. The truncated form is more rapidly hydrolyzed through proteasome-mediated degradation
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-
?
topoisomerase II alpha + H2O
?
show the reaction diagram
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cleavage of topoisomerase II alpha by granzyme M at Leu1280 separates topoisomerase II alpha functional domains from the nuclear localization signals and leads to nuclear exit of topoisomerase II alpha catalytic activity
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?
tumor necrosis factor receptor-associated protein 1 + H2O
?
show the reaction diagram
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-
-
-
?
additional information
?
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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
alpha-Tubulin + H2O
?
show the reaction diagram
-
direct alpha-tubulin proteolysis by granzyme M is complex and occurs at multiple cleavage sites, one of them being Leu at position 269. Granzyme M disturbs tubulin polymerization dynamics in vitro and induces microtubule network disorganization in tumor cells in vivo
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-
?
ezrin + H2O
?
show the reaction diagram
-
granzyme M directly and efficiently cleaves the actin-plasma membrane linker ezrin
-
-
?
heterogeneous nuclear ribonucleoprotein K + H2O
?
show the reaction diagram
-
-
granzyme M most efficiently cleaves heterogeneous nuclear ribonucleoprotein K in the presence of RNA at multiple sites, thereby likely destroying heterogeneous nuclear ribonucleoprotein K function
-
?
nucleophosmin + H2O
?
show the reaction diagram
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the nucleolar phosphoprotein, nucleophosmin (NPM), is cleaved and inactivated. Targeting of NPM by granzyme M may contribute to tumor cell eradication by abolishing NPM function
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-
?
Protein + H2O
?
show the reaction diagram
additional information
?
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
in Gzm M there are cationic sites, cs1 and cs2, that may participate in binding of Gzm M to the cell surface, thereby promoting its uptake and eventual release into the cytoplasm
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
alpha1-Antichymotrypsin
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-
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Alpha1-proteinase inhibitor
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proteinase inhibitor 9
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-
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additional information
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
survivin silencing promotes X-linked inhibitor of apoptosis protein degradation and enhances GzmM-induced caspase activation as well as GzmM- and natural killer cell-induced cytolysis of target tumor cells
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.8
acetyl-KEPL-7-amido-4-methylcoumarin
-
-
2.4
acetyl-KEPM-7-amido-4-methylcoumarin
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-
0.72
acetyl-KVAM-7-amido-4-methylcoumarin
-
-
0.37
acetyl-KVPL-7-amido-4-methylcoumarin
-
-
1.3
acetyl-KVPM-7-amido-4-methylcoumarin
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-
0.4
acetyl-KYAL-7-amido-4-methylcoumarin
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-
1
acetyl-KYPL-7-amido-4-methylcoumarin
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-
2
acetyl-KYPM-7-amido-4-methylcoumarin
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-
1.8
acetyl-MEPL-7-amido-4-methylcoumarin
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-
1.5
acetyl-RYPL-7-amido-4-methylcoumarin
-
-
1.3
acetyl-VPL-7-amido-4-methylcoumarin
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-
0.0366 - 0.0423
N-benzyloxycarbonyl-(N-(beta-t-butylseryl))-prolyl-methionine-thiobenzylester
0.0263 - 0.083
N-benzyloxycarbonyl-(N-(epsilon-t-butyloxycarbonyllysyl))-prolyl-methionine-thiobenzylester
0.0334 - 0.129
N-benzyloxycarbonyl-(N-(gamma-t-butylaspartyl))-prolyl-methionine-thiobenzylester
0.079 - 0.635
N-benzyloxycarbonyl-alanyl-alanyl-methionine-thiobenzylester
0.188 - 1.02
N-benzyloxycarbonyl-alanyl-alanyl-prolyl-methionine-thiobenzylester
0.267 - 0.695
N-benzyloxycarbonyl-alanyl-aspartyl-methionine-thiobenzylester
0.793 - 3.89
N-benzyloxycarbonyl-alanyl-lysyl-methionine-thiobenzylester
0.22 - 0.694
N-benzyloxycarbonyl-alanyl-prolyl-methionine-thiobenzylester
0.21 - 2.66
N-benzyloxycarbonyl-alanyl-seryl-methionine-thiobenzylester
0.418 - 0.441
N-benzyloxycarbonyl-aspartyl-prolyl-methionine-thiobenzylester
0.361 - 0.894
N-benzyloxycarbonyl-lysyl-prolyl-methionine-thiobenzylester
0.036 - 0.181
N-benzyloxycarbonyl-phenylalanyl-prolyl-methionine-thiobenzylester
0.0915 - 0.217
N-benzyloxycarbonyl-prolyl-prolyl-methionine-thiobenzylester
0.92 - 6.65
N-benzyloxycarbonyl-seryl-prolyl-methionine-thiobenzylester
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.79
acetyl-KEPL-7-amido-4-methylcoumarin
Homo sapiens
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-
0.42
acetyl-KEPM-7-amido-4-methylcoumarin
Homo sapiens
-
-
0.077
acetyl-KVAM-7-amido-4-methylcoumarin
Homo sapiens
-
-
0.7
acetyl-KVPL-7-amido-4-methylcoumarin
Homo sapiens
-
-
0.74
acetyl-KVPM-7-amido-4-methylcoumarin
Homo sapiens
-
-
0.06
acetyl-KYAL-7-amido-4-methylcoumarin
Homo sapiens
-
-
0.77
acetyl-KYPL-7-amido-4-methylcoumarin
Homo sapiens
-
-
0.115
acetyl-KYPM-7-amido-4-methylcoumarin
Homo sapiens
-
-
0.142
acetyl-MEPL-7-amido-4-methylcoumarin
Homo sapiens
-
-
0.22
acetyl-RYPL-7-amido-4-methylcoumarin
Homo sapiens
-
-
0.0044
acetyl-VPL-7-amido-4-methylcoumarin
Homo sapiens
-
-
0.0917 - 0.161
N-benzyloxycarbonyl-(N-(beta-t-butylseryl))-prolyl-methionine-thiobenzylester
0.03 - 0.335
N-benzyloxycarbonyl-(N-(epsilon-t-butyloxycarbonyllysyl))-prolyl-methionine-thiobenzylester
0.042 - 2.13
N-benzyloxycarbonyl-(N-(gamma-t-butylaspartyl))-prolyl-methionine-thiobenzylester
2.46 - 10.8
N-benzyloxycarbonyl-alanyl-alanyl-methionine-thiobenzylester
13.1 - 30
N-benzyloxycarbonyl-alanyl-alanyl-prolyl-methionine-thiobenzylester
0.0903 - 0.438
N-benzyloxycarbonyl-alanyl-aspartyl-methionine-thiobenzylester
6.07 - 9.68
N-benzyloxycarbonyl-alanyl-lysyl-methionine-thiobenzylester
10.6 - 18.1
N-benzyloxycarbonyl-alanyl-prolyl-methionine-thiobenzylester
1.68 - 12.8
N-benzyloxycarbonyl-alanyl-seryl-methionine-thiobenzylester
17.3 - 29
N-benzyloxycarbonyl-aspartyl-prolyl-methionine-thiobenzylester
0.031 - 13.1
N-benzyloxycarbonyl-lysyl-prolyl-methionine-thiobenzylester
0.0784 - 6.08
N-benzyloxycarbonyl-phenylalanyl-prolyl-methionine-thiobenzylester
0.00785 - 6.08
N-benzyloxycarbonyl-prolyl-prolyl-methionine-thiobenzylester
0.26 - 255
N-benzyloxycarbonyl-seryl-prolyl-methionine-thiobenzylester
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
32000 - 35000
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SDS-PAGE, glycosylated forms
33000 - 83000
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SDS-PAGE, differentially glycosylated species in Pichia pastoris
35000
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SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
proteolytic modification
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first step requires cleavage of signal peptide resulting in an inactive zymogen with an amino-terminal activation propeptide, which must then be cleaved by another protease to generate a biologically active enzyme
additional information
-
post-translational modification, including O-glycosylation of serine, phosphorylation of serine and threonine and myristoylation of glycine, play an important role in the interaction of enzyme with the cell surface membrane and regulate protein trafficking and stability. Phosphorylated serine and threonine also plays a role in tumor elimination, viral clearance and tissue repair
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structures of wild-type hGzmM, the inactive D86N-GzmM mutant with bound peptide substrate, and the complexes with a catalytic product and with a tetrapeptide chloromethylketone inhibitor are solved to 1.96 A, 2.30 A, 2.17 A and 2.70 A, respectively
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, Hepes buffer, pH 7.5, several months
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by cation exchange chromatography, more than 98% purified
-
by cation-exchange chromatography
-
by Ni-affinity chromatography
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by Ni-affinity chromatography; nickel affinity column chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
D86N-GzmM cDNA inserted into the Gal4 DNA binding domain vector. pGBKT7-D86N-GzmM and pGADT7-survivin cotransformed into the yeast stain AH109. Active GzmM and inactive D86N-GzmM mutant expressed in Escherichia coli Rosetta (DE3) strain. Stable HeLa transfectants or Jurkat cells incubated with different concentrations of GzmM or mutant D86N-GzmM
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expressed in Pichia pastoris
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expressed in Pichia pastoris; expressed in Pichia pastoris strain X33
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expression in Pichia pastoris
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into transfer vector pBacPAK8, expressed in Sf21 cells
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recombinant GrzM produced from Pichia pastoris
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subcloned from I.M.A.G.E. clone 112558 into vector pPiCza A for expression in Pichia pastoris
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wild-type, mutant with deletion of a dipeptide and mutant with deletion of a hexapeptide, expressed in COS cells
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
CD4+ T cells hardly express GrM (only 4%). GrM protein expression is absent in non-T non-natural killer cells in peripheral blood mononuclear cells
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granzyme M is expressed by lymphocytes of both the innate and adaptive immune system: high expression by natural killer cells, natural killer T cells, and gammadelta T cells (70-80% of cells). CD8+ T cells also express GrM (ca. 50% of cells) and comparing the naive to early effector-memory, to late effectormemory, to effector subset, this expression gradually increases during differentiation (in particular by the differentiated effector CD27- CD45RO- subset). GrM is usually co-expressed with perforin (ca. 60% of cells). In CD8+ T cells, about 90% of GrM positive cells also express GrB
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GrM protein expression by lymphocyte populations is not significantly affected by a panel of GrB-inducing cytokines
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GrzM is exclusively expressed by natural killer cells
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K179M
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reduced activity for substrate containing methionine at P1, acquired chymase activity for phenylalanine at P1
S182A
-
inactive
S195A
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inactive
S201G
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reduced activity for substrate containing methionine at P1, acquired chymase activity for phenylalanine at P1
additional information
-
GrzM-deficient mice, which display altered murine CMV pathology and normal recovery from ectromelia virus infection
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine