Information on EC 3.4.16.6 - carboxypeptidase D

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The expected taxonomic range for this enzyme is: Eukaryota

EC NUMBER
COMMENTARY
3.4.16.6
-
RECOMMENDED NAME
GeneOntology No.
carboxypeptidase D
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
preferential release of a C-terminal arginine or lysine residue
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
-
-
hydrolysis of peptide bond
-
-
hydrolysis of peptide bond
-
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
carboxypeptidase Kex1
-
-
-
-
cereal serine carboxypeptidase II
-
-
-
-
CP-MII.1
-
-
-
-
CP-MII.2
-
-
-
-
CP-MII.3
-
-
-
-
CP-WII
-
-
-
-
CPD
-
-
-
-
CPDW-II
-
-
-
-
EC 3.4.12.1
-
-
formerly
-
EC 3.4.16.1
-
-
formerly
-
EC 3.4.21.13
-
-
formerly
-
gene KEX1 serine carboxypeptidase
-
-
-
-
Kex-1 endopeptidase
-
-
Kex-1 protease
-
-
KEX1 carboxypeptidase
-
-
-
-
Kex1 protease
-
-
KEX1 proteinase
-
-
-
-
Saccharomyces cerevisiae KEX1 gene product
-
-
-
-
serine carboxypeptidase B-like protease
-
-
wheat carboxypeptidase II
-
-
CAS REGISTRY NUMBER
COMMENTARY
153967-26-1
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
enzyme encoded by silver gene svr
-
-
Manually annotated by BRENDA team
cytokine-inducible isoform CPD-N with incomplete N-terminal domain and intact domains II and III
-
-
Manually annotated by BRENDA team
expression in BSC-40 cells
-
-
Manually annotated by BRENDA team
KEX1 gene from Saccharomyces cerevisiae expressed using the baculovirus/insect cell system
-
-
Manually annotated by BRENDA team
truncated KEX1 gene expressed in the baculovirus/insect cell system
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
alpha-factor-Lys-Arg + H2O
mature active alpha-factor + Lys + Arg
show the reaction diagram
-
maturation takes place in sequential manner
-
?
benzoyl-Phe-Ala-Arg + H2O
benzoyl-Phe-Ala + Arg
show the reaction diagram
-
-
-
-
?
dansyl-L-Ala-L-Arg + H2O
dansyl-L-Ala + L-Arg
show the reaction diagram
-
-
-
-
?
Dansyl-Phe-Ala-Arg + H2O
Dansyl-Phe-Ala + Arg
show the reaction diagram
-
-
-
-
?
furylacryloyl-Ala-Arg + H2O
furylacryloyl-Ala + Arg
show the reaction diagram
-
-
-
-
?
furylacryloyl-Ala-Arg + H2O
furylacryloyl-Ala + Arg
show the reaction diagram
-
activity assay
-
-
?
[Leu5]enkephalin-Arg6 + H2O
?
show the reaction diagram
-
-
-
-
?
[Leu5]enkephalin-Lys6 + H2O
?
show the reaction diagram
-
-
-
-
?
[Met5]enkephalin-Arg6 + H2O
?
show the reaction diagram
-
-
-
-
?
[Met5]enkephalin-Lys6 + H2O
?
show the reaction diagram
-
-
-
-
?
furylacryloyl-Ala-Lys + H2O
furylacryloyl-Ala + Lys
show the reaction diagram
-
-
-
?
additional information
?
-
-
specificity
-
-
-
additional information
?
-
P09620
specificity
-
-
-
additional information
?
-
-
specificity
-
-
-
additional information
?
-
-
activity of individual domains of the enzyme. Domain 1B is more active at neutral pH and greatly prefers C-terminal Arg over Lys, whereas domain 2 is more active at pH 5-6 and slightly prefers C-terminal Lys over Arg
-
-
-
additional information
?
-
P09620
the enzyme has a narrow specificity for lysyl ar arginyl residues at the C-terminus of the substrate
-
-
-
additional information
?
-
-
role in the processing of both the alpha-pheromone and the killer toxin precursors
-
-
-
additional information
?
-
-
involved in the C-terminal processing of the lysine and arginine residues from the precursors of K1 and K2 killer toxins and alpha-factor (mating pheromone)
-
-
-
additional information
?
-
-
the enzyme functions in processing of proteins that transit the secretory pathway
-
-
-
additional information
?
-
-
plays a role in polypeptide processing in yeast
-
-
-
additional information
?
-
-
specific for the removal of basic amino acids from prohormone processing intermediates, in mammalian cells
-
-
-
additional information
?
-
-
preferential release of a C-terminal arginine or lysine residue
-
-
-
additional information
?
-
-
preferential release of a C-terminal arginine or lysine residue
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
additional information
?
-
-
role in the processing of both the alpha-pheromone and the killer toxin precursors
-
-
-
additional information
?
-
-
involved in the C-terminal processing of the lysine and arginine residues from the precursors of K1 and K2 killer toxins and alpha-factor (mating pheromone)
-
-
-
additional information
?
-
-
the enzyme functions in processing of proteins that transit the secretory pathway
-
-
-
additional information
?
-
-
plays a role in polypeptide processing in yeast
-
-
-
additional information
?
-
-
specific for the removal of basic amino acids from prohormone processing intermediates, in mammalian cells
-
-
-
additional information
?
-
-
preferential release of a C-terminal arginine or lysine residue
-
-
-
additional information
?
-
-
preferential release of a C-terminal arginine or lysine residue
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
activates at low concentrations
Mg2+
-
activates at low concentrations
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1,10-phenanthroline
-
noncompetitive
1-chloro-3-tosylamido-7-amino-2-heptanone
-
-
alpha-Factor KR
-
-
-
chymostatin
-
pH 5.6, 50% inhibition at 13000 nM, pH 6.5, 50% inhibition at 2200 nM. Comparison with inhibitory effect on other peptidases
diisopropyl fluorophosphate
-
-
DL-2-mercaptomethyl-3-guanidinoethylthiopropanoic acid
-
competitive
ebelactone B
-
pH 5.6, 50% inhibition at 2100 nM, pH 6.5, 50% inhibition at 1500 nM. Comparison with inhibitory effect on other peptidases
Enkephalin heptapeptides
-
-
Guanidinoethylmercaptosuccinic acid
-
-
Guanidinoethylmercaptosuccinic acid
-
domain 1B and domain
lactacystin
-
pH 5.6, 50% inhibition at 1400 nM, pH 6.5, 50% inhibition at 4300 nM. Comparison with inhibitory effect on other peptidases
omuralide
-
i.e. (1S,4S,5R)-1-[(1S)-1-hydroxy-2-methylpropyl]-4-methyl-6-oxa-2-azabicyclo[3.2.0]heptane-3,7-dione, pH 5.6, 50% inhibition at 4.8 nM, pH 6.5, 50% inhibition at 2 nM. Comparison with inhibitory effect on other peptidases
PCMB
-
domain 1B and domain
phenylmethylsulfonyl fluoride
-
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.284
-
Benzoyl-Phe-Ala-Arg
-
-
0.335
-
Benzoyl-Phe-Ala-Arg
-
-
0.063
-
dansyl-L-Ala-L-Arg
-
pH 5.6, 37C
0.132
-
dansyl-L-Ala-L-Arg
-
pH 5.6, 37C
0.208
-
dansyl-Phe-Ala-Arg
-
pH 7.4, 37C, domain 1B
0.246
-
dansyl-Phe-Ala-Arg
-
pH 5.7, 37C, domain 2
0.516
-
furylacryloyl-Ala-Arg
-
-
0.962
-
furylacryloyl-Ala-Lys
-
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
3.86
-
dansyl-Phe-Ala-Arg
-
pH 5.7, 37C, domain 2
32.6
-
dansyl-Phe-Ala-Arg
-
pH 7.4, 37C, domain 1B
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
60 U/ml, purified truncated Kex-1-C611
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
7
-
wild-type enzyme
5.6
-
-
domain 1B
5.6
-
-
and 6.5-7.0. Activity maximum at pH 5.6 is due to activity of domain II, at pH 6.5-7.0 due to domain I
6.3
-
-
activity assay
6.5
7
-
and 5.6. Activity maximum at pH 5.6 is due to activity of domain II, at pH 6.5-7.0 due to domain I
7.4
-
-
domain 2
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
-
cleavage assay
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
pI-value below pH 3.0
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
exclusive expression of 180 kDa isoform
Manually annotated by BRENDA team
-
T-lymphoma cell, exclusive synthesis of isoform SPD-N
Manually annotated by BRENDA team
-
lymphoma cell, exclusive synthesis of isoform SPD-N
Manually annotated by BRENDA team
-
Schneider 2 cell line, each splice variant of enzyme occurs in this cell line. Short enzyme forms containing a single carboxypeptidase domain are secreted from S2 cells, while long forms containing three carboxypeptidase domains, a transmembrane domain, and one out of two cytosolic domains are retained. C-terminal tail 2 leads to Golgi localization. The two C-terminal tails result in different internalization efficiencies from the cell surface
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
mainly trans-Golgi network
Manually annotated by BRENDA team
-
enzyme isoforms bearing C-terminal tail sequence 2
Manually annotated by BRENDA team
-
98% of enzymic activity
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
58000
-
-
calculated, truncated Kex-1-C611
67000
-
-
determined by SDS-PAGE, truncated Kex-1-C611
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 54219, calculation from nucleotide sequence
additional information
-
enzyme co-immunoprecipitates with protein phosphatase 2A and alpha4 phosphoprotein
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
glycoprotein
-
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
crystal structure of KEX1DELTAp, an enzyme form lacking the acidic and the membrane-spanning domain at 2.4 A resolution; structure of Kex1DELTAp, an enzyme that lacks the acidic domain and membrane-spanning portion of Kex1p
P09620
crystal structure at 2.2 A resolution
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
domain 1B and 2 expressed in Sf9 cells using baculovirus expression system
-
archieved by a two-step chromatographic process using cation-exchange, SP Sepharose, followed by anion-exchange, DEAE Sepharose FF resin
-
KEX1 gene from Saccharomyces cerevisiae expressed using the baculovirus/insect cell system
-
using IgG-Sepharose
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
individual domains of the enzyme are expressed in insect Sf9 cells using the baculovirus expression system. Medium from domain 1B-expressing cells and domain 2-expressing cells shows substantial enzymatic activity, whereas medium from domain 1A-expressing cells is not different from cells infected with wild-type virus. The individual domains 1A, 1B, and 2 are expressed in baculovirus under the polyhedrin promoter and with the signal peptide of the rat enzyme
-
a truncated form of Kex-1, Kex-1-C611, is constructed comprising the amino acid residues from 104-611 and lacks the transmembrane domain
-
into the yeast shuttle vector pVT100-ZZ
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
E353Q
-
cells infected with the mutant enzyme show considerable carboxypeptidase activity, mutation eliminates the activity of domain 1
E771Q
-
cells infected with the mutant enzyme show considerable carboxypeptidase activity, mutation eliminates the activity of domain 2
additional information
-
enzyme is encoded by silver gene svr. Mutant lines svrPG33 do not survive past the early larval stage, having a P-element insertion upstream of the initiation ATG. Mutant lines svr1 and svrpoi are viable, with silvery body color and poited wings. svr1 gene has a three-nucleotide deletion in exon 6, svrpoi has a 1072-bp duplication of the gene that introduces a stop codon into the open reading frame. Both mutations eliminate enzyme activity of the second carboxypeptidase-like domain, which reduces levels of the long forms of enzyme but does not affect the levels of the short forms
additional information
-
studies of dependence of Saccharomyces cerevisiae K2 preprotoxin killing and immunity properties modulated by the action of Kex1p and Kex2p enzymes show that the lack of Kex1p carboxypeptidase 10times decreases toxin activity, and the deficiency of Kex2p endopeptidase completely removes killing ability
additional information
-
lack of the Kex1 protease results in fusion defects during yeast mating
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
biotechnology
-
Kex-1-C611 is proposed as a convenient biotechnological reagent for the cleavage of fusion proteins
molecular biology
-
Kex2 and Kex1 may promote cell fusion by proteolytically processing substrates that act in parallel to Prm1 (pheromone-regulated membrane protein 1) as an alternative fusion machine, as cell wall components, or both