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Information on EC 3.4.17.12 - carboxypeptidase M and Organism(s) Homo sapiens and UniProt Accession P14384

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EC Tree
     3 Hydrolases
         3.4 Acting on peptide bonds (peptidases)
             3.4.17 Metallocarboxypeptidases
                3.4.17.12 carboxypeptidase M
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This record set is specific for:
Homo sapiens
UNIPROT: P14384 not found.
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Word Map
The taxonomic range for the selected organisms is: Homo sapiens
The enzyme appears in selected viruses and cellular organisms
Synonyms
cpm, carboxypeptidase m, carboxypeptidase-m, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
carboxypeptidase M
-
carboxypeptidase-M
-
-
M14006 (Merops-ID)
-
-
-
-
additional information
-
the enzyme belongs to the CPN subgroup of mammalian carboxypeptidases
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
cleavage of C-terminal arginine or lysine residues from polypeptides
show the reaction diagram
catalytic mechanism, active site groove structure involving residues Y242, W241, K228, K269, E264, R127, H66, E69, and H173, substrate interactins
cleavage of C-terminal arginine or lysine residues from polypeptides
show the reaction diagram
substrate binding via residue Tyr266
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
hydrolysis of peptide bond
CAS REGISTRY NUMBER
COMMENTARY hide
120038-28-0
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
Ala-Ser-His-Leu-Gly-Leu-Ala-Arg + H2O
Ala-Ser-His-Leu-Gly-Leu-Ala + Arg
show the reaction diagram
human anaphylatoxin C3A fragment 70-77
-
-
?
benzoyl-Ala-Arg + H2O
benzoyl-Ala + Arg
show the reaction diagram
-
-
-
?
benzoyl-Asn-Arg + H2O
benzoyl-Asn + Arg
show the reaction diagram
-
-
-
?
benzoyl-Gln-Arg + H2O
benzoyl-Gln + Arg
show the reaction diagram
-
-
-
?
benzoyl-Gly-Arg + H2O
benzoyl-Gly + Arg
show the reaction diagram
-
-
-
?
benzoyl-Gly-argininic acid + H2O
?
show the reaction diagram
-
-
-
?
benzoyl-Gly-argininic acid + H2O
benzoyl-Gly + argininic acid
show the reaction diagram
-
-
-
?
benzoyl-Gly-L-Arg + H2O
benzoyl-Gly + L-Arg
show the reaction diagram
-
-
-
?
benzoyl-Gly-Lys + H2O
benzoyl-Gly + Lys
show the reaction diagram
-
-
-
?
benzoyl-His-Arg + H2O
benzoyl-His + Arg
show the reaction diagram
-
-
-
?
benzoyl-Ile-Arg + H2O
benzoyl-Ile + Arg
show the reaction diagram
-
-
-
?
benzoyl-L-Asn-L-Arg + H2O
benzoyl-L-Asn + L-Arg
show the reaction diagram
-
-
-
?
benzoyl-Leu-Arg + H2O
benzoyl-Leu + Arg
show the reaction diagram
-
-
-
?
benzoyl-Met-Arg + H2O
benzoyl-Met + Arg
show the reaction diagram
-
-
-
?
benzoyl-Phe-Arg + H2O
benzoyl-Phe + Arg
show the reaction diagram
-
-
-
?
benzoyl-Ser-Arg + H2O
benzoyl-Ser + Arg
show the reaction diagram
-
-
-
?
benzoyl-Thr-Arg + H2O
benzoyl-Thr + Arg
show the reaction diagram
-
-
-
?
benzoyl-Trp-Arg + H2O
benzoyl-Trp + Arg
show the reaction diagram
-
-
-
?
benzoyl-Tyr-Arg + H2O
benzoyl-Tyr + Arg
show the reaction diagram
-
-
-
?
benzoyl-Val-Arg + H2O
benzoyl-Val + Arg
show the reaction diagram
-
-
-
?
bradykinin(-Phe-Arg) + H2O
?
show the reaction diagram
-
-
-
?
Dansyl-Ala-Arg + H2O
Dansyl-Ala + Arg
show the reaction diagram
enzyme assay
-
-
?
dansyl-L-Ala-L-Arg + H2O
dansyl-L-Ala + L-Arg
show the reaction diagram
-
-
-
?
furylacryloyl-Ala-Arg + H2O
furylacryloyl-Ala + Arg
show the reaction diagram
-
-
-
?
furylacryloyl-Ala-Lys + H2O
furylacryloyl-Ala + Lys
show the reaction diagram
-
-
-
?
Gly-Gly-Arg + H2O
Gly-Gly + Arg
show the reaction diagram
-
-
-
?
Gly-Leu-Ala-Arg + H2O
Gly-Leu-Ala + Arg
show the reaction diagram
human anaphylatoxin C3A fragment 74-77
-
-
?
Hippuryl-L-Lys + H2O
Hippuric acid + L-Lys
show the reaction diagram
-
-
-
?
His-Leu-Gly-Leu-Ala-Arg + H2O
His-Leu-Gly-Leu-Ala + Arg
show the reaction diagram
human anaphylatoxin C3A fragment 72-77
-
-
?
Leu-Ala-Arg + H2O
Leu-Ala + Arg
show the reaction diagram
human anaphylatoxin C3A fragment 75-77
-
-
?
Leu-Gly-Leu-Ala-Arg + H2O
Leu-Gly-Leu-Ala + Arg
show the reaction diagram
human anaphylatoxin C3A fragment 73-77
-
-
?
N-(4-hydroxybenzoyl)-Gly-Arg + H2O
N-(4-hydroxybenzoyl)-Gly + L-arginine
show the reaction diagram
-
-
-
?
Pro-Gly-Lys-Ala-Arg + H2O
Pro-Gly-Lys-Ala + Arg
show the reaction diagram
-
-
-
?
Ser-His-Leu-Gly-Leu-Ala-Arg + H2O
Ser-His-Leu-Gly-Leu-Ala + Arg
show the reaction diagram
human anaphylatoxin C3A fragment 71-77
-
-
?
Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg + H2O
Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe + arginine
show the reaction diagram
-
bradykinin
-
-
?
benzoyl-Ala-Lys + H2O
benzoyl-Ala + Lys
show the reaction diagram
-
-
-
?
benzoyl-Gly-Arg + H2O
benzoyl-Gly + Arg
show the reaction diagram
benzoyl-Gly-Lys + H2O
benzoyl-Gly + Lys
show the reaction diagram
bradykinin + H2O
?
show the reaction diagram
-
-
-
-
?
dansyl-Ala-Arg + H2O
?
show the reaction diagram
-
substrate for determination of CPM activity
-
-
?
Dansyl-Ala-Arg + H2O
Dansyl-Ala + Arg
show the reaction diagram
dynorphin A(1-13) + H2O
Tyr-Gly-Gly-Phe-Leu-Arg-Arg-Ile-Arg-Pro-Lys-Leu + Lys
show the reaction diagram
-
i.e. Tyr-Gly-Gly-Phe-Leu-Arg-Arg-Ile-Arg-Pro-Lys-Leu-Lys
-
-
?
EGF + H2O
des-Arg53-EGF + arginine
show the reaction diagram
-
EGF, epidermal growth factor
-
-
?
epidermal growth factor + H2O
dea-Arg53-epidermal growth factor + arginine
show the reaction diagram
-
-
-
-
?
furylacryloyl-L-Ala-L-Arg + H2O
furylacryloyl-L-Ala + L-Arg
show the reaction diagram
-
-
-
-
?
Leu5-Arg6-enkephalin + H2O
?
show the reaction diagram
-
-
-
-
?
Lys-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg + H2O
Lys-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe + arginine
show the reaction diagram
-
kallidin
-
-
?
Met5-Arg6-enkephalin + H2O
?
show the reaction diagram
-
-
-
-
?
Met5-Lys6-enkephalin + H2O
?
show the reaction diagram
-
-
-
-
?
SDF-1alpha + H2O
des-lys SDF-1alpha + lysine
show the reaction diagram
-
stromal cell-derived factor
-
-
?
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
EGF + H2O
des-Arg53-EGF + arginine
show the reaction diagram
-
EGF, epidermal growth factor
-
-
?
epidermal growth factor + H2O
dea-Arg53-epidermal growth factor + arginine
show the reaction diagram
-
-
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Zn2+
the catalytic center is located in the catalytic domain in a cavity with a funnel-like access that contains the catalytic Zn2+ ion
KCl
-
addition of 0.5 M NaCl to the assay buffer does not significantly alter the activities of the wilde-type enzyme with 0.2 mM dansyl-Ala-Arg or the E260Q mutant enzyme but does substantially increase the activity of the E260A mutant, 219%. The enhancement of E260A is not specific for NaCl, as similar increases are detected with other salts such as NaNO3, 249%, KNO3, 256%, KCl, 256%, or Na2SO4, 297%
KNO3
-
addition of 0.5 M NaCl to the assay buffer does not significantly alter the activities of the wilde-type enzyme with 0.2 mM dansyl-Ala-Arg or the E260Q mutant enzyme but does substantially increase the activity of the E260A mutant, 219%. The enhancement of E260A is not specific for NaCl, as similar increases are detected with other salts such as NaNO3, 249%, KNO3, 256%, KCl, 256%, or Na2SO4, 297%
Na2SO4
-
addition of 0.5 M NaCl to the assay buffer does not significantly alter the activities of the wilde-type enzyme with 0.2 mM dansyl-Ala-Arg or the E260Q mutant enzyme but does substantially increase the activity of the E260A mutant, 219%. The enhancement of E260A is not specific for NaCl, as similar increases are detected with other salts such as NaNO3, 249%, KNO3, 256%, KCl, 256%, or Na2SO4, 297%
NaCl
-
addition of 0.5 M NaCl to the assay buffer does not significantly alter the activities of the wilde-type enzyme with 0.2 mM dansyl-Ala-Arg or the E260Q mutant enzyme but does substantially increase the activity of the E260A mutant, 219%. The enhancement of E260A is not specific for NaCl, as similar increases are detected with other salts such as NaNO3, 249%, KNO3, 256%, KCl, 256%, or Na2SO4, 297%
NaNO3
-
addition of 0.5 M NaCl to the assay buffer does not significantly alter the activities of the wilde-type enzyme with 0.2 mM dansyl-Ala-Arg or the E260Q mutant enzyme but does substantially increase the activity of the E260A mutant, 219%. The enhancement of E260A is not specific for NaCl, as similar increases are detected with other salts such as NaNO3, 249%, KNO3, 256%, KCl, 256%, or Na2SO4, 297%
additional information
-
the enzyme is a metallopeptidase
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
-
1,4-dithiothreitol
-
2-Guanidinoethylmercaptosuccinic acid
strong CPM inhibitor
2-mercaptomethyl-3-guanidinoethyl thiopropanoic acid
strong CPM inhibitor
1,10-phenanthroline
2-mercaptomethyl-3-guanidinoethyl thiopropanoic acid
-
-
2-mercaptomethyl-3-guanidinoethylthiopropanoic acid
-
-
Cadmium acetate
-
-
Cd(CH3COO)2
-
-
DL-2-mercaptomethyl-3-guanidino-ethylthiopropanoic acid
-
-
DL-2-mercaptomethyl-3-guanidinoethylthiopropanoic acid
-
inhibits best at acidic pH
DL-mercaptomethyl-3-guanidino-ethylthiopropanoic acid
-
-
Guanidinoethyl mercaptosuccinic acid
Guanidinoethylmercaptosuccinic acid
-
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
G-CSF
-
granulocyte-colony stimulating factor
-
IFN-gamma
-
CPM activity can be upregulated approximately two-fold in HLMVECs by IL-1beta plus IFN-gamma treatment
-
IL-1beta
-
CPM activity can be upregulated approximately two-fold in HLMVECs by IL-1beta plus IFN-gamma treatment, IL-1beta alone can upregulate CPM expression
-
lipopolysaccharide
-
following stimulation with bacterial lipopolysaccharide a marginal upregulation of the expression of carboxypeptidase M is observed
additional information
-
prolactin/17beta-estradiol does not increase carboxypeptidase M expression
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.033
Ala-Ser-His-Leu-Gly-Leu-Ala-Arg
CPMwt, pH 7.4, 37°C
0.115 - 0.18
benzoyl-Ala-Arg
0.343
benzoyl-Asn-Arg
CPMwt, pH 7.4, 37°C
0.127
benzoyl-Gln-Arg
CPMwt, pH 7.4, 37°C
0.255 - 0.29
benzoyl-Gly-Arg
0.046 - 0.049
benzoyl-Gly-argininic acid
10.15 - 10.65
Benzoyl-Gly-Lys
0.239
benzoyl-His-Arg
CPMwt, pH 7.4, 37°C
0.205
benzoyl-Ile-Arg
CPMwt, pH 7.4, 37°C
0.168
benzoyl-Leu-Arg
CPMwt, pH 7.4, 37°C
0.13 - 0.138
benzoyl-Met-Arg
0.0584
benzoyl-Phe-Arg
CPMwt, pH 7.4, 37°C
0.219
benzoyl-Ser-Arg
CPMwt, pH 7.4, 37°C
0.192
benzoyl-Thr-Arg
CPMwt, pH 7.4, 37°C
0.069
benzoyl-Trp-Arg
CPMwt, pH 7.4, 37°C
0.048
benzoyl-Tyr-Arg
CPMwt, pH 7.4, 37°C
0.264
benzoyl-Val-Arg
CPMwt, pH 7.4, 37°C
0.0277
bradykinin(-Phe-Arg)
CPMwt, pH 7.2, 37°C
0.073 - 0.083
Dansyl-Ala-Arg
0.068 - 0.09
furylacryloyl-Ala-Arg
1.152
furylacryloyl-Ala-Lys
CPMwt, pH 7.2, 37°C
1.181
Gly-Gly-Arg
CPMwt, pH 7.2, 37°C
0.05
Gly-Leu-Ala-Arg
CPMwt, pH 7.4, 37°C
0.028
His-Leu-Gly-Leu-Ala-Arg
CPMwt, pH 7.4, 37°C
0.027
Leu-Ala-Arg
CPMwt, pH 7.4, 37°C
0.041
Leu-Gly-Leu-Ala-Arg
CPMwt, pH 7.4, 37°C
0.206 - 0.241
N-(4-hydroxybenzoyl)-Gly-Arg
0.035
Pro-Gly-Lys-Ala-Arg
CPMwt, pH 7.2, 37°C
0.051
Ser-His-Leu-Gly-Leu-Ala-Arg
CPMwt, pH 7.4, 37°C
0.016
bradykinin
0.059 - 0.28
Dansyl-Ala-Arg
0.063
Leu5-Arg6-enkephalin
-
-
0.046
Met5-Arg6-enkephalin
-
-
0.375
Met5-Lys6-enkephalin
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
7.4
Ala-Ser-His-Leu-Gly-Leu-Ala-Arg
CPMwt, pH 7.4, 37°C
77 - 107
benzoyl-Ala-Arg
35
benzoyl-Asn-Arg
CPMwt, pH 7.4, 37°C
22
benzoyl-Gln-Arg
CPMwt, pH 7.4, 37°C
3.9 - 6.07
benzoyl-Gly-Arg
273 - 316
benzoyl-Gly-argininic acid
9.6 - 11
Benzoyl-Gly-Lys
14
benzoyl-His-Arg
CPMwt, pH 7.4, 37°C
1
benzoyl-Ile-Arg
CPMwt, pH 7.4, 37°C
19
benzoyl-Leu-Arg
CPMwt, pH 7.4, 37°C
87 - 130
benzoyl-Met-Arg
33
benzoyl-Phe-Arg
CPMwt, pH 7.4, 37°C
60
benzoyl-Ser-Arg
CPMwt, pH 7.4, 37°C
22.9
benzoyl-Thr-Arg
CPMwt, pH 7.4, 37°C
23
benzoyl-Trp-Arg
CPMwt, pH 7.4, 37°C
27
benzoyl-Tyr-Arg
CPMwt, pH 7.4, 37°C
7.7
benzoyl-Val-Arg
CPMwt, pH 7.4, 37°C
30
bradykinin(-Phe-Arg)
CPMwt, pH 7.2, 37°C
17 - 17.6
Dansyl-Ala-Arg
32 - 40
furylacryloyl-Ala-Arg
11
furylacryloyl-Ala-Lys
CPMwt, pH 7.2, 37°C
1.9
Gly-Gly-Arg
CPMwt, pH 7.2, 37°C
8.2
Gly-Leu-Ala-Arg
CPMwt, pH 7.4, 37°C
6.6
His-Leu-Gly-Leu-Ala-Arg
CPMwt, pH 7.4, 37°C
4.2
Leu-Ala-Arg
CPMwt, pH 7.4, 37°C
6.1
Leu-Gly-Leu-Ala-Arg
CPMwt, pH 7.4, 37°C
4.9 - 5.7
N-(4-hydroxybenzoyl)-Gly-Arg
11
Pro-Gly-Lys-Ala-Arg
CPMwt, pH 7.2, 37°C
6.7
Ser-His-Leu-Gly-Leu-Ala-Arg
CPMwt, pH 7.4, 37°C
2.45
bradykinin
0.233 - 6.02
Dansyl-Ala-Arg
1.77
Leu5-Arg6-enkephalin
-
-
15.6
Met5-Arg6-enkephalin
-
-
11.1
Met5-Lys6-enkephalin
-
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000003 - 0.000006
MERGETPA
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
14
CPMwt with dansyl-Ala-Arg
19
hCPM with dansyl-Ala-Arg
1.344
-
-
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.2
enzyme assay
6.5 - 7.5
-
-
7 - 8
-
-
additional information
-
neutral pH-optimum
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 8
-
pH 5.5: about 35% of maximal activity, pH 7.0-8.0: activity maximum
5.5 - 9
-
pH 5.5: 25% of maximal activity, pH 9.0: about 60% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
enzyme assay
37
-
activity assay
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
very low activity
Manually annotated by BRENDA team
CPM mRNA expression is very low in the human heart
Manually annotated by BRENDA team
CPM expression is the highest in kidneys
Manually annotated by BRENDA team
CPM mRNA expression is very low in the human liver
Manually annotated by BRENDA team
CPM expression is the highest in lung membrane fractions
Manually annotated by BRENDA team
CPM expression is the highest in macrophages
Manually annotated by BRENDA team
CPM mRNA expression is very low in the human pancreas
Manually annotated by BRENDA team
very low activity
Manually annotated by BRENDA team
-
interstitial trophoblast
Manually annotated by BRENDA team
-
erythroid cell progenitor
Manually annotated by BRENDA team
-
myeloid cell progenitor
Manually annotated by BRENDA team
-
megakaryocytic cell progenitor
Manually annotated by BRENDA team
-
peripheral
Manually annotated by BRENDA team
-
mobilized peripheral blood CD34+ cell
Manually annotated by BRENDA team
-
tumor cells of renal cell carcinoma subtypes lose carboxypeptidase M expression upon dedifferentiation. There is a correlation between low carboxypeptidase M expression and tumor cell type. Carboxypeptidase M staining is intense on phagocytotic tumor-associated macrophages, and enzyme is also detected in the tumor-associated vasculature. Coexistence of carboxypeptidase M and the epidermal growth factor receptor is detected in papillary renal cell carcinoma
Manually annotated by BRENDA team
-
endovascular
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
anchored via glycosylphosphatidylinositol
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
CBPM_HUMAN
443
0
50514
Swiss-Prot
Secretory Pathway (Reliability: 1)
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
48000
deglycosylated protein, calculated from amino acid sequence
49000
determined by SDS-PAGE
50000
1 * 50000, recombinant 3-truncated, glycosylphosphatidylinositol-free enzyme, SDS-PAGE
47600
-
1 * 62000, glycosylated enzyme, SDS-PAGE, 1 * 47600, deglycosylated enzyme, SDS-PAGE
50000
-
x * 50000, SDS-PAGE
62000
73000
-
gel filtration
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
monomer
-
1 * 62000, glycosylated enzyme, SDS-PAGE, 1 * 47600, deglycosylated enzyme, SDS-PAGE
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
glycoprotein
additional information
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified, recombinant, glycosylphosphatidylinositol-free enzyme, X-ray diffraction structure determination and analysis at 3.0 A resolution, modelling
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E264Q
the mutation completely eliminates the CPM activity
E260A
-
mutation reduces the ratio of turnover-numver to Km-value by 104fold and further decreases stability.Addition of 0.5 M NaCl to the assay buffer does not significantly alter the activities of the wild-type enzyme with 0.2 mM dansyl-Ala-Arg or the E260Q mutant enzyme but does substantially increase the activity of the E260A mutant, 219%. The enhancement of E260A is not specific for NaCl, as similar increases are detected with other salts such as NaNO3, 249%, KNO3, 256%, KCl, 256%, or Na2SO4, 297%
E260Q
-
mutation has minimal effects on kinetic parameters, but decreased heat stability
E264Q
-
mutation results in a 10000fold decrease in activity, but the enzyme still binds to p-aminobenzoylarginine-Sepharose and is resistant to trypsin treatment, indicating that the protein is folded properly
S406A
-
very similar to the wild-type enzyme with regard to expression and release by phosphatidylinositol-specific phospholipase C
S406P
-
the wild-type and S406A and S406T mutants are expressed on the plasma membrane in glycosylphosphatidylinositol-anchored form, the S406P mutant is not and is retained in a perinuclear location
S406T
-
very similar to the wild-type enzyme with regard to expression and release by phosphatidylinositol-specific phospholipase C
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
1 h, room temperature, 33% loss of activity
647252
4.3
-
1 h, room temperature, 17% loss of activity
647252
4.5 - 5
-
1 h, room temperature, stable
647252
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
wild-type enzyme and mutant enzymes E260Q and E260A are stable
45
-
15 min, 50% inactivation of mutant enzyme E260A
49
-
15 min, 15 min, 50% inactivation of mutant enzyme E260Q
50
-
15 min, complete inactivation of mutant enzyme E260A
53
-
15 min, wild-type enzyme retains 50% of its activity
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 50% v/v glycerol, stable for at least 1 year
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant 3'-truncated, glycosylphosphatidylinositol-free, soluble enzyme from insect cells by anion exchange chromatography and gel filtration to homogeneity
wild type carboxypeptidase M, CPMwt, is purified from Pichia pastoris supernatant using hydrophobic interaction chromatography and affinity chromatography, hCPM is purified from human prostasomes
native enzyme from placental microvillus membranes by solubilization through either phosphatidylinositol-specific phospholipase C or CHAPS, ion exchange and arginine affinity chromatography, and gel filtration
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recombinant enzyme 1753fold from Pichia pastoris strain GS115 by gel filtration and anion exchange chromatography to homogeneity
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the CPM secreted in the supernatant is purified by a two-step procedure consisting of gel filtration and ion-exchange chromatography
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in HEK-293 cells, in Pichia pastoris, and in baculovirus-infected insect cells
expression of 3'-truncated, glycosylphosphatidylinositol-free, soluble enzyme in insect cells using the baculovirus transfection system
placental cDNA is incorporated in the pICZalphaA shuttle vector, signal peptide and C-terminal hydrophobic membrane anchor signal are removed
CPM cDNAs are cloned into pcDNA3 or pcDNA6 for expression in mammalian cells, to express N-terminal CFP- or YFP-tagged CPM pECFP-C1 and pEYFP-C1 vectors are used, to generate a fusion protein of CPM attached to the extracellular N-terminus of B1R pcDNA3 and 6 vectors are used
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DNA and amino acid sequence determination and analysis, genetic structure, localization on chromosome 12q13-q15, expression in insect cells via baculovirus transfection
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expression in Pichia pastoris
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expression of wild-type and mutant enzymes in COS-7 cells or HEK-293 cells. The wild-type and S406A and S406T mutants are expressed on the plasma membrane in glycosylphosphatidylinositol-anchored form, the S406P mutant is notz and is retained in a perinuclear location. Expression in baculovirus infected cells in a glycophosphatidyl-anchored form, whereas a truncated form, lacking the putative signal sequence for glycosylphosphatidyl anchoring is secreted at high levels into the medium. Both forms have lower molecular masses than native placental enzyme indicating a minimal glycosylation
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PCR-amplified fragment encoding hCPM is cloned into the pGEM-T-Easy cloning vector, and subsequent into the pPIC9 vector for transfomation of Pichia pastoris cells
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
oxidative stress reduction by epoetin delta goes along with the upregulation of renoprotective genes. The expression of carboxypeptidase M is significantly upregulated upon erythropoietin administration in and only in those cell cultures in which an erythropoietin-induced cytoprotective effect is seen. In cultures in which epoetin delta does not show protective effects, no upregulation of carboxypeptidase M is observed
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
diagnostics
expression is positively correlated with overall survival and negatively correlated with recurrence, lymph node invasion, and N stage in colorectal cancer
medicine
diagnostics
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CPM, a gene that encodes carboxypeptidase M, is consistently amplified in liposarcomas but not in different subtypes of lipoma or normal fat, CPM could be used as an alternative and novel diagnostic tool for these tumors
medicine
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Deddish, P.A.; Skidgel, R.A.; Erds, E.G.
Enhanced Co2+ activation and inhibitor binding of carboxypeptidase M at low pH. Similarity to carboxypeptidase H (enkephalin convertase)
Biochem. J.
261
289-291
1989
Homo sapiens
Manually annotated by BRENDA team
Skidgel, R.A.; Davis, R.M.; Tan, F.
Human carboxypeptidase M. Purification and characterization of a membrane-bound carboxypeptidase that cleaves peptide hormones
J. Biol. Chem.
264
2236-2241
1989
Homo sapiens
Manually annotated by BRENDA team
Tan, F.; Deddish, P.A.; Skidgel, R.A.
Human carboxypeptidase M
Methods Enzymol.
248
663-675
1995
Homo sapiens
Manually annotated by BRENDA team
Tan, F.; Balsitis, S.; Black, J.K.; Blochl, A.; Mao, J.F.; Becker, R.P.; Schacht, D.; Skidgel, R.A.
Effect of mutation of two critical glutamic acid residues on the activity and stability of human carboxypeptidase M and characterization of its signal for glycosylphosphatidylinositol anchoring
Biochem. J.
370
567-578
2003
Homo sapiens
Manually annotated by BRENDA team
Craveiro, R.B.; Ramalho, J.D.; Chagas, J.R.; Wang, P.H.; Casarini, D.; Pesquero, J.L.; Araujo, R.C.; Pesquero, J.B.
High expression of human carboxypeptidase M in Pichia pastoris: purification and partial characterization
Braz. J. Med. Biol. Res.
39
211-217
2006
Homo sapiens
Manually annotated by BRENDA team
Skidgel, R.A.
Carboxypeptidase M
Handbook of Proteolytic Enzymes (Barrett, J. ; Rawlings, N. D. ; Woessner, J. F. , eds. ) Academic Press
2
851-854
2004
Canis lupus familiaris, Homo sapiens
-
Manually annotated by BRENDA team
Reverter, D.; Maskos, K.; Tan, F.; Skidgel, R.A.; Bode, W.
Crystal structure of human carboxypeptidase M, a membrane-bound enzyme that regulates peptide hormone activity
J. Mol. Biol.
338
257-269
2004
Homo sapiens (P14384), Homo sapiens
Manually annotated by BRENDA team
Matthews, K.W.; Mueller-Ortiz, S.L.; Wetsel, R.A.
Carboxypeptidase N: a pleiotropic regulator of inflammation
Mol. Immunol.
40
785-793
2004
Homo sapiens
Manually annotated by BRENDA team
Deiteren, K.; Surpateanu, G.; Gilany, K.; Willemse, J.L.; Hendriks, D.F.; Augustyns, K.; Laroche, Y.; Scharpe, S.; Lambeir, A.M.
The role of the S1 binding site of carboxypeptidase M in substrate specificity and turn-over
Biochim. Biophys. Acta
1774
267-277
2007
Homo sapiens (P14384)
Manually annotated by BRENDA team
Skidgel, R.A.; Stanisavljevic, S.; Erdoes, E.G.
Kinin- and angiotensin-converting enzyme (ACE) inhibitor-mediated nitric oxide production in endothelial cells
Biol. Chem.
387
159-165
2006
Bos taurus, Homo sapiens
Manually annotated by BRENDA team
Schremmer-Danninger, E.; Naegler, D.K.; Miska, K.; Flaig, M.J.; Faussner, A.; Fink, E.; Raggi, M.C.; Jochum, M.; Rehbock, J.
Kinin receptors in stimulated and characterized decidua tissue-derived cells
Int. Immunopharmacol.
7
103-112
2007
Homo sapiens
Manually annotated by BRENDA team
Zhang, X.; Tan, F.; Zhang, Y.; Skidgel, R.A.
Carboxypeptidase M and kinin B1 receptors interact to facilitate efficient B1 signaling from B2 agonists
J. Biol. Chem.
283
7994-8004
2008
Homo sapiens
Manually annotated by BRENDA team
Tsakiris, I.; Soos, G.; Nemes, Z.; Kiss, S.S.; Andras, C.; Szanto, J.; Dezso, B.
The presence of carboxypeptidase-M in tumour cells signifies epidermal growth factor receptor expression in lung adenocarcinomas: The coexistence predicts a poor prognosis regardless of EGFR levels
J. Cancer Res. Clin. Oncol.
134
439-451
2008
Homo sapiens
Manually annotated by BRENDA team
Fujiwara, H.
Membrane-bound peptidases regulate human extravillous trophoblast invasion
Placenta
28 Suppl A
S70-S75
2007
Homo sapiens
Manually annotated by BRENDA team
Marquez-Curtis, L.; Jalili, A.; Deiteren, K.; Shirvaikar, N.; Lambeir, A.M.; Janowska-Wieczorek, A.
Carboxypeptidase M expressed by human bone marrow cells cleaves the C-terminal lysine of SDF-1{alpha}: Another player in hematopoietic stem/progenitor cell mobilization?
Stem Cells
26
1211-1220
2008
Homo sapiens
Manually annotated by BRENDA team
Deiteren, K.; Hendriks, D.; Scharpe, S.; Lambeir, A.M.
Carboxypeptidase M: Multiple alliances and unknown partners
Clin. Chim. Acta
399
24-39
2009
Homo sapiens (P14384), Homo sapiens
Manually annotated by BRENDA team
Abdelmagid, S.A.; Too, C.K.
Prolactin and estrogen up-regulate carboxypeptidase-D to promote nitric oxide production and survival of MCF-7 breast cancer cells
Endocrinology
149
4821-4828
2008
Homo sapiens
Manually annotated by BRENDA team
De Beuf, A.; Hou, X.H.; DHaese, P.C.; Verhulst, A.
Epoetin delta reduces oxidative stress in primary human renal tubular cells
J. Biomed. Biotechnol.
2010
395785
2010
Homo sapiens
Manually annotated by BRENDA team
Erickson-Johnson, M.R.; Seys, A.R.; Roth, C.W.; King, A.A.; Hulshizer, R.L.; Wang, X.; Asmann, Y.W.; Lloyd, R.V.; Jacob, E.K.; Oliveira, A.M.
Carboxypeptidase M: a biomarker for the discrimination of well-differentiated liposarcoma from lipoma
Mod. Pathol.
22
1541-1547
2009
Homo sapiens
Manually annotated by BRENDA team
Zhang, X.; Tan, F.; Skidgel, R.A.
Carboxypeptidase M is a positive allosteric modulator of the kinin B1 receptor
J. Biol. Chem.
288
33226-33240
2013
Homo sapiens (P14384), Homo sapiens
Manually annotated by BRENDA team
Denis, C.J.; Van Acker, N.; De Schepper, S.; De Bie, M.; Andries, L.; Fransen, E.; Hendriks, D.; Kockx, M.M.; Lambeir, A.M.
Mapping of carboxypeptidase m in normal human kidney and renal cell carcinoma: expression in tumor-associated neovasculature and macrophages
J. Histochem. Cytochem.
61
218-235
2013
Homo sapiens
Manually annotated by BRENDA team
Chu, M.; Wu, S.; Wang, W.; Yu, Y.; Zhang, M.; Sang, L.; Tian, T.; Lu, Y.; Yuan, W.; Huang, Q.; Yi, M.; Gao, Y.; Xiao, J.; Lian, Y.; Zhuang, X.; Zhang, Z.F.; Wu, J.
Functional variant of the carboxypeptidase M (CPM) gene may affect silica-related pneumoconiosis susceptibility by its expression a multistage case-control study
Occup. Environ. Med.
76
169-174
2019
Homo sapiens (P14384)
Manually annotated by BRENDA team
Lu, D.; Yao, Q.; Zhan, C.; Le-Meng, Z.; Liu, H.; Cai, Y.; Tu, C.; Li, X.; Zou, Y.; Zhang, S.
MicroRNA-146a promote cell migration and invasion in human colorectal cancer via carboxypeptidase M/src-FAK pathway
Oncotarget
8
22674-22684
2017
Homo sapiens (P14384), Homo sapiens
Manually annotated by BRENDA team