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3.4.17.12: carboxypeptidase M

This is an abbreviated version!
For detailed information about carboxypeptidase M, go to the full flat file.

Word Map on EC 3.4.17.12

Reaction

cleavage of C-terminal arginine or lysine residues from polypeptides =

Synonyms

carboxypeptidase M, carboxypeptidase-M, CPM, M14006 (Merops-ID), More

ECTree

     3 Hydrolases
         3.4 Acting on peptide bonds (peptidases)
             3.4.17 Metallocarboxypeptidases
                3.4.17.12 carboxypeptidase M

Cloned

Cloned on EC 3.4.17.12 - carboxypeptidase M

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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
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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expressed in Escherichia coli strain DH5alpha
expressed in HEK-293 cells, in Pichia pastoris, and in baculovirus-infected insect cells
expression in Pichia pastoris
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expression of 3'-truncated, glycosylphosphatidylinositol-free, soluble enzyme in insect cells using the baculovirus transfection system
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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into the pCMV5, pGex4T-1 and pMalC2 vector for expression in COS-7 and Escherichia coli cells
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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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placental cDNA is incorporated in the pICZalphaA shuttle vector, signal peptide and C-terminal hydrophobic membrane anchor signal are removed