Information on EC 3.4.21.41 - complement subcomponent C1r

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The expected taxonomic range for this enzyme is: Tetrapoda

EC NUMBER
COMMENTARY hide
3.4.21.41
-
RECOMMENDED NAME
GeneOntology No.
complement subcomponent C1r
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Selective cleavage of Lys(or Arg)-/-Ile bond in complement subcomponent C1s to form C_overbar_1s_ (EC 3.4.21.42)
show the reaction diagram
EC 3.4.21.42
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
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endopeptidase
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CAS REGISTRY NUMBER
COMMENTARY hide
80295-69-8
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
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binding residues are conserved in the CUB1 domains of C1r, MASP-1/-3, and MASP-2, indicating that the interaction mechanism is conserved in initiating complexes of the lectin and classical pathways
metabolism
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
acetyl-Arg methyl ester + H2O
acetyl-Arg methanol
show the reaction diagram
-
-
-
-
?
acetyl-Gly-Lys-methyl ester + H2O
?
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Gly-Arg-thiobenzyl ester + H2O
?
show the reaction diagram
-
-
-
?
Benzyloxycarbonyl-Lys-thiobenzyl ester + H2O
?
show the reaction diagram
-
-
-
?
chordin + H2O
?
show the reaction diagram
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C-terminal domains CUB1 and CUB2 are required for the ventralizing activity of Xld and for its ability to cleave chordin
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-
?
complement C1q zymogen + H2O
active complement C1q + ?
show the reaction diagram
complement component C1s
?
show the reaction diagram
complement component C1s + H2O
activated complement component C1s + ?
show the reaction diagram
complement component C1s + H2O
complement component C1sbar
show the reaction diagram
MASP-3 K448Q zymogen + H2O
active MASP-3 protein + ?
show the reaction diagram
-
poor activity with the wild-type MASP-3
-
-
?
N-acetyl-Gly-Lys methyl ester + H2O
N-acetyl-Gly-Lys + methanol
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-Gly-Arg thiobenzyl ester + H2O
?
show the reaction diagram
-
-
-
-
?
N-carbobenzoxy-Lys-p-nitrophenyl ester + H2O
N-carbobenzoxy-Lys + 4-nitrophenyl
show the reaction diagram
-
-
-
-
?
N-carbobenzoxy-Tyr-p-nitrophenyl ester + H2O
N-carbobenzoxy-Tyr + 4-nitrophenol
show the reaction diagram
-
-
-
-
?
zymogen C1s + H2O
active C1s protease + ?
show the reaction diagram
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the enzyme is active with the substrate fragment consisting of complement control protein domains, CCP1 and CCP2, plus serine protease domain of wild-type and mutant Q462N, Q462G, I464A, and Q462N/I464A forms of C1s, mutant Q462N/I464A substrate gives very low activity
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-
?
zymogen C1s + H2O
active protease C1s + ?
show the reaction diagram
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-
-
-
?
zymogen C1s + H2O |
active C1s protease + ?
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
complement C1q zymogen + H2O
active complement C1q + ?
show the reaction diagram
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-
-
-
?
complement component C1s
?
show the reaction diagram
complement component C1s + H2O
activated complement component C1s + ?
show the reaction diagram
zymogen C1s + H2O
active protease C1s + ?
show the reaction diagram
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-
-
-
?
zymogen C1s + H2O |
active C1s protease + ?
show the reaction diagram
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-
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?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3-alkoxy-4-chloro-7-guanidinoisocoumarin
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4-chloro-3-[3-isothiureidopropoxyl]isocoumarin
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4-Nitrophenyl-4-guanidinobenzoate
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C1 inhibitor
C1-inhibitor
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regulates the classical and the lectin complement pathway inhibiting complement components C1r and C1s, and MASP-2, factor XIa, Factor XIIa, and plasma kallikrein, mechanism, overview
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C1bar-inhibitor
C1INH
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sole inhibitor of the activated proteases C1r and C1s. Hereditary angioedema results from functional deficiency of the C1 inhibitor (C1INH) protein, which plays a key role in the classical pathway of complement activation
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Ca2+
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reaction with C1s
calreticulin
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prevents C1 formation
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dichloroisocoumarin
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diisopropyl fluorophosphate
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Leupeptin
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competitive
NaCl
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above 200 mM
p-amidinophenylmethylsulfonyl fluoride
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p-tosyl-Lys-chloromethyl ketone
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phenylmethylsulfonyl fluoride
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polyanethol sulfonate
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additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
20
acetyl-Gly-Lys methyl ester
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0.000062
MASP-3 K448Q zymogen
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pH 7.4, 37C
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0.000022
zymogen C1s
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pH 7.4, 37C, substrate is a fragment consisting of complement control protein domains, CCP1 and CCP2, plus serine
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additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0021
MASP-3 K448Q zymogen
Homo sapiens
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pH 7.4, 37C
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18.3
N-acetyl-Arg methyl ester
Homo sapiens
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12.5
N-Acetyl-Gly-Lys methyl ester
Homo sapiens
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0.0645
zymogen C1s
Homo sapiens
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pH 7.4, 37C, substrate is a fragment consisting of complement control protein domains, CCP1 and CCP2, plus serine
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additional information
additional information
Homo sapiens
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activated C1r and its wild-type CCP1/2-SP and CCP2-SP fragments all cleave acetyl-Gly-Lys-methyl ester with turnover numbers ranging from 8.2 s to 9.2 s
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
340
MASP-3 K448Q zymogen
Homo sapiens
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pH 7.4, 37C
202404
2900
zymogen C1s
Homo sapiens
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pH 7.4, 37C, substrate is a fragment consisting of complement control protein domains, CCP1 and CCP2, plus serine
202403
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.4
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assay at
7.5 - 8.5
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
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assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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nasopharyngeal carcinoma cells
Manually annotated by BRENDA team
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MEF-2 cell, low-density lipoprotein receptor-related protein 1-deficient macrophages show increased expression of C1r
Manually annotated by BRENDA team
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C1r mRNA expression is significantly increased in RAW 264.7 cells in which low-density lipoprotein receptorrelated protein 1 is silenced
Manually annotated by BRENDA team
additional information
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PEA-10 cell and B-41 cell
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
16600
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gel filtration
90000
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immunoblot analysis, in PEA-10 and B-41 cells, single-chain form, which is the zymogen
790000
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additional information
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 88000, C1r zymogen, domain structure, overview
heteropentamer
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C1q, 2 * C1s, 2 * C1r
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
proteolytic modification
side-chain modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure analysis, PDB IDs 1APQ
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hanging drop vapour diffusion method, crystal structure of two fragments from the human C1r catalytic domain, each encompassing the second complement control protein CCP2 module and the SP domain. The wild-type species has an active structure, whereas the S637A mutant is a zymogen
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hanging drop vapour diffusion, crystal structure of a mutated proenzyme form of the catalytic domain of human C1r, and the serine protease domain solved and refined to 2.9 A resolution
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purified recombinant isolated active catalytic region forming a dimer in a head-to-tail fashion, complex of enzyme and product, hanging drop vapour diffusion method, 15C, mixing of 0.008 ml protein solution, containing 0.2 mg/ml protein, 20 mM Tris-HCl, pH 7.4, and 130 mM NaC, with 0.008 ml reservoir solution containing 14% w/v PEG 6000, 0.2 M NaCl, 10% v/v glycerol, and 0.1 M Tris-HCl, pH 7.4, cryoprotection of crystals in 20% glycerol, X-ray diffraction structure determination and analysis at 2.6 A resolution, molecular replacement
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
after reduction and alkylation C1rbar undergoes proteolytic cleavage which lead to the successive removal of two fragments of 35000 Da and of 7000-10000 Da, leaving a divalent molecule of reduced size. The product C1rbar II retains the original antigenic properties of C1rbar and a functional active site, but loses the capacity to bind C1s
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C1rbar undergoes autolytic cleavage
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
2C, pH 5.3-7.3, in absence of Ca2+, 2 weeks, stable
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
commercial preparation
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from human plasma
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from inclusion bodies, ion exchange chromatography (Q-Sepharose), gel filtration
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proenzyme C1r
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recombinant active catalytic region of C1r from Escherichia coli strain BL21(DE3) by ion exchange chromatography and gel filtration
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solubilization of inclusion-bodies
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
baculovirus-mediated expression is used to produce fragments containing the SP domain and either 2 CCP modules (CCP1/2-SP) or only the second CCP module (CCP2-SP). In each case the wild-type species and two mutants stabilized in the proenzyme form by mutations at the cleavage site (R446Q) or at the active site serine residue (S637A) are produced
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CUB2-complement control protein 1 module expressed in Escherichia coli BL21(DE3) pLysS
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development and evaluation of a method for expression of recombinant full-length human C-terminally tagged C1q involving stable transfection of HEK 293-F mammalian cells
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expression in insect cells
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expression in insect Sf9,High5 cells
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expression of complement control protein module 2 (I356-V433), complement control protein module 1 - complement control protein module 2 (I289-V433), CUB2 - complement control protein module 1 (Q173-D358) and CUB2 - complement control protein module 1 - complement control protein module 2 (Q173-V433)
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expression of modules: Q173-D358 (CUB2-complement control protein 1) and I356-V433 (complement control protein 2)
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expression of the isolated active catalytic region, containing the C-terminal serine protease domain, and two preceding complement control protein modules, of C1r in Escherichia coli strain BL21(DE3)
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production of the CCP1-CCP2-SP, the CCP2-SP and the SP fragments in Escherichia coli
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recombinant expression of full-length C1r protease and the CUB1-EGF-CUB2 fragment with low yield
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Xld is overexpressed in Xenopus embryos
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
R446Q
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stabilized in the single-chain proenzyme form by mutation at the cleavage site, no esterolytic activity with acetyl-Gly-Lys-methyl ester
S637A
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stabilized in the single-chain proenzyme form by mutation at the active site serine residue, no esterolytic activity with acetyl-Gly-Lys-methyl ester
S654A
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mutant without autoactivation
additional information
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by the point mutation of C1r - Arg-Phe - of the natural cleavage site Arg-Ile - the zymogen is stabilized, while the biological activity is not affected
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
reconstitution of the C1 complex using purified C1q, C1r and C1s in the presence of C1-INH, interaction with C-reactive protein CRP, overview
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
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anti-inflammatory functions are exerted via inhibition of complement system proteases (C1r, C1s, MASP2) and the plasma kallikrein-kinin system proteases