Information on EC 3.4.21.61 - Kexin

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
3.4.21.61
-
RECOMMENDED NAME
GeneOntology No.
Kexin
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Cleavage of -Lys-Arg-/- and -Arg-Arg-/- bonds to process yeast alpha-factor pheromone and killer toxin precursors
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
99676-46-7
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
overproducing strain
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
-
an interaction between the prodomain and C-terminal domain regulates the secretion of PCSK9
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
Ac-(beta-cyclohexyl)alanineYKK 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + Ac-(beta-cyclohexyl)alanineYKK
show the reaction diagram
-
-
-
?
Ac-Ala-Tyr-Lys-Arg 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + ?
show the reaction diagram
-
-
-
?
Ac-Ala-Tyr-Lys-Lys 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + ?
show the reaction diagram
-
-
-
?
Ac-alpha-aminobutyric acid-YKK 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + Ac-alpha-aminobutyric acid-YKK
show the reaction diagram
-
-
-
?
Ac-Arg-Tyr-Lys-Lys 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + Ac-Arg-Tyr-Lys-Lys
show the reaction diagram
-
-
-
?
Ac-AYKK 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + Ac-AYKK
show the reaction diagram
-
-
-
?
Ac-AYKR 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + Ac-AYKR
show the reaction diagram
-
-
-
?
Ac-Cit-Tyr-Lys-Lys 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + Ac-Cit-Tyr-Lys-Lys 4-methylcoumarin
show the reaction diagram
-
this substrate is cleaved poorly
-
?
Ac-CYKK 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + Ac-CYKK
show the reaction diagram
-
-
-
?
Ac-FYKK 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + Ac-FYKK
show the reaction diagram
-
-
-
?
Ac-Leu-Lys-Arg-p-nitroanilide + H2O
?
show the reaction diagram
-
-
-
?
Ac-Nle-Tyr-Lys-Arg 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + Ac-Nle-Tyr-Lys-Arg-COOH
show the reaction diagram
Ac-Nle-Tyr-Lys-Lys 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + ?
show the reaction diagram
-
-
-
?
Ac-Nle-YKK 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + Ac-Nle-YKK
show the reaction diagram
-
-
-
?
Ac-Nle-YKR 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + ?
show the reaction diagram
-
-
-
?
Ac-Nle-YKR 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + Ac-Nle-YKR
show the reaction diagram
-
-
-
?
Ac-norvaline-YKK 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + Ac-norvaline-YKK
show the reaction diagram
-
-
-
?
Ac-Pro-Met-Tyr-Lys-Arg 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + ?
show the reaction diagram
-
-
-
?
Ac-Pro-Met-Tyr-Lys-Arg 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + Ac-Pro-Met-Tyr-Lys-Arg
show the reaction diagram
-
-
-
?
Ac-RYKK 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + Ac-RYKK
show the reaction diagram
-
-
-
?
Ac-VYKK 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + Ac-VYKK
show the reaction diagram
-
-
-
?
alpha-mating factor + H2O
?
show the reaction diagram
-
cleavage of dibasic sites
-
-
-
Arg-Lys(DABCYL)-Nle-Tyr-Lys-Arg-Glu-Ala-Glu-Ala-Glu(EDANS)-Arg + H2O
Arg-Lys(DABCYL)-Nle-Tyr-Lys-Arg + Glu-Ala-Glu-Ala-Glu(EDANS)-Arg
show the reaction diagram
-
-
-
?
Arg-Lys(DABCYL)-Nle-Tyr-Lys-Lys-Glu-Ala-Glu-Ala-Glu(EDANS)-Arg + H2O
Arg-Lys(DABCYL)-Nle-Tyr-Lys-Lys + Glu-Ala-Glu-Ala-Glu(EDANS)-Arg
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Ala-Tyr-Lys-Lys 4-methylcoumarin 7-amide + H2O
7-hydroxy-4-methylcoumarin + benzyloxycarbonyl-Ala-Tyr-Lys-Lys
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Nle-Tyr-Lys-Arg 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Nle-Tyr-Lys-Lys 4-methylcoumarin 7-amide + H2O
7-hydroxy-4-methylcoumarin + Boc-Nle-Tyr-Lys-Lys
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Nle-YKR 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
-
-
?
Benzyloxycarbonyl-Tyr-Lys-Arg 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
CLC chloride channel + H2O
proteolytically processed CLC chloride channel
show the reaction diagram
-
cleavage in first intracellular loop at residues K136/R137
-
-
?
D-Ac-Nle-Tyr-Lys-Arg 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
-
-
?
Dpy-5 procollagen + H2O
?
show the reaction diagram
-
-
-
-
?
Killer toxin precursors + H2O
?
show the reaction diagram
-
cleavage of dibasic sites
-
-
-
L-pyroglutamyl-Arg-Thr-Lys-Arg-4-methylcoumaryl-7-amide + H2O
L-pyroglutamyl-Arg-Thr-Lys-Arg + 7-amino-4-methyl-coumarin
show the reaction diagram
-
-
-
-
?
low density lipoprotein receptor
?
show the reaction diagram
-
-
-
-
?
low density lipoprotein receptor + H2O
?
show the reaction diagram
low density lipoprotein receptor-related protein 1 + H2O
?
show the reaction diagram
low-density lipoprotein receptor
?
show the reaction diagram
PCSK9 posttranslationally promotes the degradation of the low-density lipoprotein receptor
-
-
?
N-tert-butyloxycarbonyl-Gly-Lys-Arg 4-methylcoumarin 7-amide + H2O
N-tert-butyloxycarbonyl-Gly-Lys-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
59.2% of the activity with tert-butyloxycarbonyl-Gln-Arg-Arg 4-methylcoumarin 7-amide
-
-
-
pGlu-Arg-Thr-Lys-Arg-4-methylcoumaryl-7-amide + H2O
7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
Precursor protein of the mating hormone alpha-factor of Saccharomyces cerevisiae + H2O
?
show the reaction diagram
prekallikrein
?
show the reaction diagram
-
cleaved into four fragments by ASP, protein cleaved at specific sequences
-
-
?
Prm2
?
show the reaction diagram
-
-
-
-
?
pro-alpha-mating factor + H2O
alpha-mating factor + ?
show the reaction diagram
proGIP + H2O
GIP
show the reaction diagram
Proinsulin + H2O
Insulin + ?
show the reaction diagram
-
cleaves human proinsulin at the peptide bond between Arg32 and Glu33
-
?
proopiomelanocortin + H2O
beta-lipotropic hormone + adrenocorticotropic hormone + ?
show the reaction diagram
-
-
-
-
?
Protein + H2O
?
show the reaction diagram
t-butyloxycarbonyl-Arg-Val-Arg-Arg-4-methylcoumaryl-7-amide + H2O
t-butyloxycarbonyl-Arg-Val-Arg-Arg + 7-amino-4-methyl-coumarin
show the reaction diagram
-
-
-
-
?
t-butyloxycarbonyl-EKK 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + t-butyloxycarbonyl-EKK
show the reaction diagram
-
-
-
?
t-butyloxycarbonyl-Glu-Lys-Lys-4-methylcoumaryl-7-amide + H2O
t-butyloxycarbonyl-Glu-Lys-Lys + 7-amino-4-methyl-coumarin
show the reaction diagram
-
best substrate
-
-
?
t-butyloxycarbonyl-Gly-Lys-Arg-4-methylcoumaryl-7-amide + H2O
t-butyloxycarbonyl-Gly-Lys-Arg + 7-amino-4-methyl-coumarin
show the reaction diagram
-
-
-
-
?
t-butyloxycarbonyl-QGR 4-methylcoumarin 7-amide + H2O
7-amino-4-methylcoumarin + t-butyloxycarbonyl-QGR
show the reaction diagram
-
-
-
?
tert-Butyloxycarbonyl-Ala-Pro-Arg 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
17% of the activity with tert-butyloxycarbonyl-Gln-Arg-Arg 4-methylcoumarin 7-amide
-
-
-
tert-butyloxycarbonyl-Gln-Arg-Arg 4-methylcoumarin 7-amide + H2O
tert-butyloxycarbonyl-Gln-Arg-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
-
tert-Butyloxycarbonyl-Leu-Arg-Arg 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
116% of the activity with tert-butyloxycarbonyl-Gln-Arg-Arg 4-methylcoumarin 7-amide
-
-
-
tert-Butyloxycarbonyl-Leu-Lys-Arg 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
92.8% of the activity with tert-butyloxycarbonyl-Gln-Arg-Arg 4-methylcoumarin 7-amide
-
-
-
tert-Butyloxycarbonyl-Val-Pro-Arg 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
38% of the activity with tert-butyloxycarbonyl-Gln-Arg-Arg 4-methylcoumarin 7-amide
-
-
-
Ykl077w
?
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
alpha-mating factor + H2O
?
show the reaction diagram
-
cleavage of dibasic sites
-
-
-
CLC chloride channel + H2O
proteolytically processed CLC chloride channel
show the reaction diagram
-
cleavage in first intracellular loop at residues K136/R137
-
-
?
Killer toxin precursors + H2O
?
show the reaction diagram
-
cleavage of dibasic sites
-
-
-
low density lipoprotein receptor + H2O
?
show the reaction diagram
low density lipoprotein receptor-related protein 1 + H2O
?
show the reaction diagram
Precursor protein of the mating hormone alpha-factor of Saccharomyces cerevisiae + H2O
?
show the reaction diagram
pro-alpha-mating factor + H2O
alpha-mating factor + ?
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
no effect with Mg2+, Mn2+, Co3+, and Fe3+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(3S,4S,5S,6R)-2-([(2R,3S,4S,5S,6S)-3,5-dihydroxy-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-4-yl]amino)-6-(hydroxymethyl)tetrahydro-2H-thiopyran-3,4,5-triol
-
i.e. BJ-12-26-1, greatly reduces the processing of substrate proopiomelanocortin
(3S,4S,5S,6R)-2-([(2S,3S,4R,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-methoxytetrahydro-2H-pyran-3-yl]amino)-6-(hydroxymethyl)tetrahydro-2H-thiopyran-3,4,5-triol
-
i.e. BJ-12-21-2, greatly reduces the processing of substrate proopiomelanocortin
Ac-R-E-R-K-chloromethylketone
-
alternate binding site and resultant displacement of the scissile bond in the active site results in a decrease in the acylation rate
Ala-Lys-Arg boronic acid
-
Ala-Lys-Arg chloromethyl ketone
-
-
CT-peptide
-
decreases PC1/3 activity at high concentrations (micromol)
-
D-Tyr-Ala-Lys-Arg-CH2Cl
-
-
D-Tyr-Pro-Gly-Lys-Arg-CH2Cl
-
-
Decanoyl-Arg-Ala-Lys-Arg-CH2Cl
-
-
Decanoyl-Arg-Glu(OMe)-Lys-Arg-CH2Cl
-
-
Decanoyl-Phe-Ala-Lys-Arg-CH2Cl
-
-
decanoyl-R-V-K-R-chloromethylketone
-
-
decanoyl-R-V-R-K-chloromethylketone
-
-
diisopropyl fluorophosphate
-
-
eglin c mutant D42R
-
-
-
eglin c mutant L45R
-
-
-
eglin c mutant Tyr replaced with Glu at P4
-
-
-
eglin variant M1 RVTR
-
-
-
eglin variant M2 RVKR
-
-
-
eglin variant M3 RVTRDERY
-
-
-
eglin variant M4 RVTRDRRY
-
-
-
eglin variant M5 RVTRDLDY
-
-
-
eglin variant M6 RVTRDLRR
-
-
-
eglin variant M7 RVTRDLRE
-
-
-
eglin variant M8 RVTRDARY
-
-
-
human urine trypsin inhibitor
-
most potent inhibitor
-
ISIS 394814
-
most potent antisense oligonucleotide. Administration to high fat-fed mice for 6 weeks reduces hepatic PCSK9 mRNA levels by 92%, total cholesterol and LDL by 53% and 38%, respectively. Inhibition of PCSK9 expression results in a 2fold increase in hepatic LDLR protein levels
-
Mercurials
-
-
-
p-chloromercuribenzoate
-
-
Peptidyl chloromethanes
-
-
Phe-Ala-Lys-Arg-CH2Cl
-
-
phenylmethylsulfonyl fluoride
Pro-norvaline-Tyr-Lys-Arg-CH2Cl
-
-
propeptide
-
addition of purified recombinant propeptide (20 nanomol) to PC1/3 enzymatic reaction leads to a 50% reduction in enzymatic activity. In the presence of 5 nanomol CT-peptide and 20 nanomol propeptide, inhibition is reduced to 35%
-
sterol
-
completely blocks increase in PCSK9 mRNA by lovastatin
TIMP-3
-
completely abolishes the production of the 34-kDa PCSK9 product
-
Tyr-Ala-Arg-Ala-Lys-Arg-CH2Cl
-
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
atorvastatin
-
significantly increases circulating PCSK9 levels by 34% compared with baseline and placebo and decreases low density lipoprotein cholesterol levels by 42%
CT-peptide
-
increases PC1/3 activity at low concentrations (nanomol). PC1/3, through its various domains, is capable of controlling its enzymatic activity in all regions of the cell that it encounters. Activation of PC1/3 by 5 nanomol CT-peptide is the same in the presence or absence of propeptide
-
K+
-
mechanism of activation
lovastatin
-
suppresses endogenous synthesis of sterols, increases both the proteins and mRNAs for PCSK9. 10 mM can reverse the increase of PCSK9 expression by lovastatin
sodium mevalonate
-
suppresses endogenous synthesis of sterols, PCSK9 is further increased
sterol-regulatory element binding protein-1
-
dramatically increases the promoter activity of PCSK9
-
sterol-regulatory element binding protein-2
-
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.35
Ac-AYKK 4-methylcoumarin 7-amide
-
pH 7.0, 21C
0.001
Ac-Nle-Tyr-Lys-Arg 4-methylcoumarin 7-amide
-
pH 7.0, 37C, in natural abundance H2O
0.038
Ac-Nle-YKK 4-methylcoumarin 7-amide
-
pH 7.0, 21C
0.001
Ac-Nle-YKR 4-methylcoumarin 7-amide
-
pH 7.0, 21C
0.023
benzyloxycarbonyl-Ala-Tyr-Lys-Lys 4-methylcoumarin 7-amide
-
pH 7.0, 21C
0.001
D-Ac-Nle-Tyr-Lys-Arg 4-methylcoumarin 7-amide
-
pH 7.0, 37C, in natural abundance H2O
0.32
t-butyloxycarbonyl-QGR 4-methylcoumarin 7-amide
-
pH 7.0, 21C
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
21.6
Ac-AYKK 4-methylcoumarin 7-amide
Saccharomyces cerevisiae
-
pH 7.0, 21C
2700
Ac-Nle-Tyr-Lys-Arg 4-methylcoumarin 7-amide
Saccharomyces cerevisiae
-
pH 7.0, 37C
300
Ac-Nle-YKK 4-methylcoumarin 7-amide
Saccharomyces cerevisiae
-
pH 7.0, 21C
3000
Ac-Nle-YKR 4-methylcoumarin 7-amide
Saccharomyces cerevisiae
-
pH 7.0, 21C
2640
benzyloxycarbonyl-Ala-Tyr-Lys-Lys 4-methylcoumarin 7-amide
Saccharomyces cerevisiae
-
pH 7.0, 21C
2940
D-Ac-Nle-Tyr-Lys-Arg 4-methylcoumarin 7-amide
Saccharomyces cerevisiae
-
pH 7.0, 37C
11.4
t-butyloxycarbonyl-EKK 4-methylcoumarin 7-amide
Saccharomyces cerevisiae
-
pH 7.0, 21C
366
t-butyloxycarbonyl-QGR 4-methylcoumarin 7-amide
Saccharomyces cerevisiae
-
pH 7.0, 21C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.002
CT-peptide
-
-
-
0.00019
decanoyl-R-V-K-R-chloromethylketone
-
-
0.00845
decanoyl-R-V-R-K-chloromethylketone
-
-
0.00000013
eglin c mutant D42R, eglin c mutant L45R
-
pH 7.5, 37C
-
0.00000003
eglin c mutant Tyr replaced with Glu at P4
-
pH 7.5, 37C
-
0.000000128
eglin variant M1 RVTR
-
pH 7.5, 37C
-
0.000000038
eglin variant M2 RVKR
-
pH 7.5, 37C
-
0.000000316
eglin variant M3 RVTRDERY
-
pH 7.5, 37C
-
0.000089
eglin variant M4 RVTRDRRY
-
pH 7.5, 37C
-
0.0000043
eglin variant M5 RVTRDLDY
-
pH 7.5, 37C
-
0.0000267
eglin variant M6 RVTRDLRR
-
pH 7.5, 37C
-
0.000039
eglin variant M7 RVTRDLRE
-
pH 7.5, 37C
-
0.000000136
eglin variant M8 RVTRDARY
-
pH 7.5, 37C
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
Homo sapiens
-
at 20C in 0.05 ml of buffer containing 10 mM Hepes, pH 7.4, 150 mM NaCl, 0.1 mM CaCl2, and 0.05% (w/v) bovine serum albumin: 0.0000048 mM with 1G08 fragment antigen binding, when wild-type expressed in HEK-293 cells, 0.0000150 mM with 1G08 fragment antigen binding, when wild-type expressed in Hep-G2 cells, 0.001 mM with 1G08 fragment antigen binding, when mutant R549A , mutant R580A/R582A and mutant E607A/K609A/E612N expressed in HEK-293 cells
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5
-
tert-butyloxycarbonyl-Gln-Arg-Arg 4-methylcoumarin 7-amide
7.2
-
benzyloxycarbonyl-Tyr-Lys-Arg 4-nitroanilide
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
expresses high levels of endogenous PC1/3
Manually annotated by BRENDA team
-
mild expression of PC1 and moderate expression of PC2. PC1 and PC2 in cerebral cortex colocalizes in pyramidal and non-pyramidal neurons with carboxypeptidase-E and somatostatin
Manually annotated by BRENDA team
-
weak expression of PC1 in CA1, CA2 and CA3 regions and nerve fibers. Moderate expression of PC2 in CA1, CA2 and CA3 regions and mild expression in nerve fibers. Colocalization of PC1 and PC2 with carboxypeptidase-E and somatostatin mostly selectively and preferentially in interneurons
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
PDB
SCOP
CATH
ORGANISM
UNIPROT
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
17640
-
mass spectrometry analysis
64000
-
mature form of wild-type PCSK9, Western blot analysis
65000
-
1 * 65000, SDS-PAGE
68000
-
x * 87000, PC1, SDS-PAGE. x * 75000, proPC2, SDS-PAGE. x * 68000, mature PC2, SDS-PAGE
75000
-
x * 87000, PC1, SDS-PAGE. x * 75000, proPC2, SDS-PAGE. x * 68000, mature PC2, SDS-PAGE
75979
-
1 * 75979, purified PCSK9, gel filtration. 1* 45708, purified PCSK9DELTAC, gel filtration
87000
-
x * 87000, PC1, SDS-PAGE. x * 75000, proPC2, SDS-PAGE. x * 68000, mature PC2, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 87000, PC1, SDS-PAGE. x * 75000, proPC2, SDS-PAGE. x * 68000, mature PC2, SDS-PAGE
monomer
additional information
-
the enzyme contains a signal peptide and a prodomain followed by a catalytic protease domain, a hinge-region and a C-terminal domain
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
-
mutation of N-glycosylation site N146 results in the decreased zymogen activation of proPC1/3 and virtually inhibits its secretion. Mutation of glycosylation site N618A mutation does not significantly affect zymogen activation. The presence, but not the exact structure, of N-glycans is crucial for proPC1/3 zymogen processing
proteolytic modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystallized by hanging drop method, in complex with an Ala-Lys-Arg boronic acid inhibitor, space group P6(5)22, a = b = 113.8 A, c = 370.2 A
in complex with Ac-Arg-Glu-Lys-Arg-peptidyl boronic acid inhibitor
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Kex2 in complex with the Ac-R-E-R-K-chloromethylketone, containing a noncognate lysine at the P1 position. Secondary subsite in the S1 pocket is present, which recognizes and binds the P1 lysine in a more shallow fashion than arginine. Kex2 contains well defined subsites that have optimally arranged electrostatic charge that positions correct substrates for hydrolysis. Chemical nature of the peptidyl inhibitor has little effect on ligand positioning at the active site in the alkylated enzyme forms
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
-
37C, t1/2: 36 min in sodium acetate buffer
29424
5.5
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37C, t1/2: 5 min in Na-MES buffer
29424
6
-
37C, 18 min in Na-MES buffer
29424
6.5
-
37C, t1/2: 17 min in Na-MES buffer
29424
7
-
37C, t1/2: 250 min in Bis-Tris-HCl buffer, 6 min in Na-HEPES buffer
29424
7.5
-
37C, t1/2: 22 min in Na-HEPES buffer
29424
8
-
37C, t1/2: 5 min in Na-HEPES buffer
29424
8.5
-
37C, t1/2: 44 min in Na-Bicine buffer
29424
9
-
37C, 17 min in Na-Bicine buffer
29424
9.5
-
37C, t1/2: 2 min
29424
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
at 4C by binding to anti-FLAG M2 affinity gel
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by Ni-nitriloacetic acid and size-exclusion chromatography, more than 95% pure
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by successive column chromatographies
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gel filtration
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on Ni-NTA resin
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recombinant enzyme
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recombinant His-tagged enzyme from HEK-293T cell medium by nickel affinity chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
adenovirus construct expressing PC1/3
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cDNAs encoding full-length PCSK9 splicing variant or mutants inserted into a modified pJB02 vector with a C-terminal Flag-HIS6 or HIS6 tag, respectively. The resulting constructs used for transient transfection of HEK-293 cells
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cloned and expressed in Pichia pastoris
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cloned KEX2 gene introduced into the kex2 mutant cells and the KEX2 gene product expressed in these cells
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expressed in HEK-293 cells, COS-1 cells or Y1 cells
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expression in HEK293 cell
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expression in HEK293 cells
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expression in HEK293T and HepG2 cells
full-length and mutant PCSK9-V5-His proteins expressed in HEK-293 cells. Truncated PCSK9DELTAC (Gln-31-Ala-451) proteins with a C-terminal His tag expressed in Escherichia coli BL21 cells
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GH4 cells co-transfected with preproGIP and PC1/3
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into vector pET-21a(+)
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isolation of the structural gene
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ligated into vector pCR2.1-Topo and subcloned into vector pET-21a(+)
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overexpressed in HEK-293 cells
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recombinant expression of enzyme mutants in Hep-G2 cells
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recombinant His-tagged enzyme catalytic domain expression in HEK-293T cells and secretion to the medium
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recombinant overexpression of the human enzyme in mouse LDL-receptor null mutant cells, expression and protein profiles, overview. Overexpression of PCSK9 increases plasma ApoB levels in an LDLR-independent fashion
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recombinant PC1/3 produced using the baculovirus expression system in Sf-9 insect cells or through intracoelemic injection in insect larvae
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transformation of the single-gene deletion strain Deltakex2 with pYEX12 plasmid
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wild type and mutant D374Y-FLAG expressed in Hep-G2 cells
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wild-type PCSK9 with a V5 epitope flag inserted into pcDNA3.1. PCSK9 mutants expressed in primary hepatocytes derived from PCSK9-/- mice
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
amounts of PCSK9 mRNA and protein in Caco-2/15 cells are associated to the regulation of 3-hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein-2 that can transcriptionally activate PCSK9 via sterol-regulatory elements located in its proximal promoter region. Depletion of cholesterol content by hydroxypropyl-beta-cyclodextrin upregulates PCSK9 transcripts (20%) and protein mass (540%), in parallel with sterol regulatory element binding protein-2 protein levels
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cholesterol (0.1 mM) solubilized in albumin or micelles significantly downregulates PCSK9 gene (30%) and protein expression (50%) in Caco-2/15 cells. Cells treated with 25-hydroxycholesterol (0.05 mM) also display significant reduction in PCSK9 gene (37%) and protein (75%) expression. Addition of bile acids taurocholate and deoxycholate to the apical culture medium lowers PCSK9 gene expression (25%) and raises PCSK9 protein expression (30%), respectively, probably via the modulation of farnesoid X receptor
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cholesteryl ester transfer protein inhibitors downregulate PCSK9 and LDLR expression through decreases in SREBP2 expression in hepatocytes. Glitazones, rapamycine, berberine, and resistin cause a reduction of enzyme expression via HNF1alpha reduction. Reduction of enzyme expression can be achieved by peroxisome proliferator-activated receptor alpha and activation of sterolresponse element binding protein 2. In HepG2 cells, hyperinsulinemia decreases the enzyme expression, an effect which is also observed in post-menopausal obese women
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in healthy men 24 h hyperinsulinemia does not alter plasma the enzyme concentrations, and the enzyme expression is similar in normal, pre- and Typ2-diabetic patients
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knockdown of PCSK9 expression in immortalized human hepatocytes using specific siRNA which results in a 38% and 53% decrease in PCSK9 protein quantity, respectively in cell lysates and culture media compared with controls
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PCSK9 is expressed throughout the entire small intestine and in enterocytes. PCSK9 is expressed almost exclusively in the epithelial barrier of the duodenum and ileum, both in enterocytes and goblet cells. PCSK9 is 160% upregulated by 0.01 mM pravastatin in CaCo-2 cells after exposure for 48 h
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PCSK9 is expressed throughout the small intestine and colon, at a level which does not vary significantly along the intestinal cephalo-caudal axis, and which is similar to that in the liver
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PCSK9 levels correlate with plasma cholesterol, LDL-C, triglycerides, fasting glucose, age and body mass index. In hypercholesterolemic patients, circulating PCSK9 is higher than in healthy volunteers, and increases with increasing statin dose, and further increased when ezetimibe is added. Ezetimibe treatment of Hep-G2 (hepatocytes) and Caco-2 (enterocytes) cells causes a slight increase in PCSK9 mRNA, but no significant rise in PCSK9 protein secretion, suggesting that these transformed cells are not ideal model cell lines
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the enzyme is up-regulated during apoptosis of neurons. Statins and increase the transcription factor SREBP2, statins also the HNF1alpha expression, and fibrates increase PPAralpha, all leading to increased enzyme PCSK9 expression. Activation of enzyme expression can be mediated by activation of insulin receptors and subsequent activation of the sterolresponse element binding protein 1 and mammalian target of rapamycin pathways
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A433T
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is associated with a plasma PCSK9 concentration of 222 ng/ml
A443T
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is expressed, processed, and secreted normally, and reduces cellular LDL uptake in a concentration-dependent like the wild-type
C678X
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a loss-of-function mutation that abolishes the release of the enzyme from the endoplasmic reticulum
DeltaN218
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in COS-1 cells only apparent upon co-expression of PC5A. Untreated Y1 cells secrete PCSK9-DeltaN218, stimulation with 8-Br-cAMP increases the level and especially that of the 34-kDa PCSK9 product
E569K
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site-directed mutagenesis, the mutant shows slightly decreased ability to block LDL uptake into HepG2 cells compared to the wild-type enzyme
E607A/K609A/E612N
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mutation does not affect the overall integrity of the PCSK9 protein, shows similar cellular uptake potencies as the wild-type, impairs 1G08-PCSK9 binding
G365R
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is associated with a plasma PCSK9 concentration of 205 ng/ml
G517R
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site-directed mutagenesis, the mutant shows highly decreased ability to block LDL uptake into HepG2 cells compared to the wild-type enzyme
N425S
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is expressed, processed, and secreted normally, and reduces cellular LDL uptake in a concentration-dependent like the wild-type
Q152H
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a dominant negative mutation that restricts enzyme proteolysis and secretion independently
Q152I
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te mutation completely abrogates proteolysis in both intra- and intermolecular systems but has only a limited impact on secretion
Q152R
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the mutation completely abolished both proteolysis and secretion
Q152X
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four phenotypes of Q152X mutants, overview
Q190R
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is associated with a plasma PCSK9 concentration of 55 ng/ml
R215H
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a naturally occurring gain-of-function mutation associated with hypercholesterolemia, the mutation impairs furin-mediated enzyme cleavage
R434W
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is associated with a plasma PCSK9 concentration of 51 ng/ml
R469W
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natural mutation, cannot modify the ability of PC5A to produce the 34-kDa PCSK9 product in HEK293 cells
R46L
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is associated with a plasma PCSK9 concentration of 51-59 ng/ml
R496W
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natural mutation, cannot modify the ability of PC5A to produce the 34-kDa PCSK9 product in HEK293 cells
R53V
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is associated with a plasma PCSK9 concentration of 39 ng/ml
R549A
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mutation does not affect the overall integrity of the PCSK9 protein, shows similar cellular uptake potencies as the wild-type, impairs 1G08-PCSK9 binding
R580A/R582A
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mutation does not affect the overall integrity of the PCSK9 protein, shows similar cellular uptake potencies as the wild-type, impairs 1G08-PCSK9 binding
R659A
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site-directed mutagenesis, the mutant shows slightly decreased ability to block LDL uptake into HepG2 cells compared to the wild-type enzyme
R659E
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site-directed mutagenesis, the mutant shows slightly decreased ability to block LDL uptake into HepG2 cells compared to the wild-type enzyme
S462P
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a loss-of-function mutation that abolishes the release of the enzyme from the endoplasmic reticulum
S636R
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site-directed mutagenesis, the mutant shows slightly decreased ability to block LDL uptake into HepG2 cells compared to the wild-type enzyme
V149A
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the mutant shows intolerance for intermolecular cleavage of the enzyme, residue Val149 is critical for secretion
V610R
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site-directed mutagenesis, the mutant shows highly decreased ability to block LDL uptake into HepG2 cells compared to the wild-type enzyme
V644R
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site-directed mutagenesis, the mutant shows highly decreased ability to block LDL uptake into HepG2 cells compared to the wild-type enzyme
C678X
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a loss-of-function mutation that abolishes the release of the enzyme from the endoplasmic reticulum
N146A
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mutation results in the decreased zymogen activation of proPC1/3 and virtually inhibits its secretion
N374A
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mutant is processed and secreted at nearly the same rate and with the same apparent molecular mass as wild-type. Residue N374 does not appear to bear an N-glycan
N618A
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mutation does not significantly affect zymogen activation
S462P
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a loss-of-function mutation that abolishes the release of the enzyme from the endoplasmic reticulum
D176G/D210A/D211S
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no preference for positively charged residues at P2 position, and S2 pocket is more solvent accessible, leading to preference for MR- over LR- or FR-containing substrates
E255I
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significantly decreased recognition of P4Arg residue in a tetrapeptide substrate
T252D
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increased recognition of Arg in P4 position, 14fold higher kcat/KM ratio for Arg than for Ala at position P6
T252D/Q283E
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increased recognition of Arg in P4 position, 15fold higher kcat/KM ratio for Arg than for Ala at position P6
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
degradation
diagnostics
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plasma enzyme concentrations are predictive for 4-5 year major cardiovascular event rate, and enzyme serum concentrations correlate with cardiovascular risk
drug development
medicine
molecular biology
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role of Kex2 during cell fusion. Kex2 may promote cell fusion by proteolytically processing substrates that act in parallel to Prm1 as an alternative fusion machine, as cell wall components, or both
additional information