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3.4.22.52: calpain-1

This is an abbreviated version!
For detailed information about calpain-1, go to the full flat file.

Word Map on EC 3.4.22.52

Reaction

broad endopeptidase specificity =

Synonyms

Cal 1, calcium-activated neutral protease I, calpain 1, calpain 1-gamma, calpain 1A, calpain I, calpain small subunit, calpain-1, calpain-1 (micro-form), calpain-I, calpain1, CANP1, CAPN1, CAPN1 g.p. (Homo sapiens), CAPN2, CAPNS1, cysteine protease, EC 3.4.22.17, EC 3.4.24.5, m-CANP, micro-calpain, mit-CPN1, mito-mu-calpain, mu-calpain, muCANP, muI-II

ECTree

     3 Hydrolases
         3.4 Acting on peptide bonds (peptidases)
             3.4.22 Cysteine endopeptidases
                3.4.22.52 calpain-1

Cloned

Cloned on EC 3.4.22.52 - calpain-1

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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
a 289 bp fragment for mu-calpain is cloned into the EcoRV site of pBluescript II KS+ vector
cloning of the 21000 Da small subunit and expression in Escherichia coli BL384
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complete amino acid sequence of the large subunit is deduced from its cDNA sequence
expressed in Escherichia coli BL21 cells
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expressed in Escherichia coli BL21(DE3) cells
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expression in Escherichia coli
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expression of mutant enzyme C115S in insect cell using a baculovirus system
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large catalytic subunit and two of its mutants are expressed in Escherichia coli using the baculovirus Sf9 system, the L-muCANPDELTA3 mutant lacks domain III, mutant L-muCANPDELTA4 lacks the calmodulin-like domain IV. In Sf9 cells co-expression of the inhibitor calpastatin is necessary to prevent autolysis of the L-muCANP subunit, whereas coexpression of the regulatory subunit enhances it. Only very low levels of mRNA of the truncated form L-muCANPDELTA4 are found in bacmid-transfectred Sf9 cells, and it proves impossible to isolate this mutant using the baculovirus expression system
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small subunit and large subunit
the cDNA fragments corresponding to the domains with four consecutive E-F hand structures in the large and small subunits are inserted into an expression vector, pUC8 or pUC17. The resulting plasmids are used to transform Escherichia coli and isopropyl-1-thio-beta-D-galactoside-inducible expression is performed