Abstract |
Chymotrypsin C is a bifunctional secretory-type serine protease in pancreas; besides proteolytical activity, it also exhibits a calcium-decreasing activity in serum. In this study, we purified activated chymotrypsin C from porcine pancreas, and identified its three active forms. Active chymotrypsin C was found to be different in the length of its 13-residue activation peptide due to carboxydipeptidase (present in the pancreas) degradation or autolysis of the activated chymotrypsin C itself, resulting in the removal of several C-terminus residues from the activation peptide. After limited chymotrypsin C cleavage with endopeptidase Lys C, several purified peptides were partially sequenced, and the entire cDNA sequence for porcine chymotrypsin C was cloned. Recombinant chymotrypsinogen C was successfully expressed in Escherichia coli cells as inclusion bodies. After refolding and activation with trypsin, the comparison of the recombinant chymotrypsin C with the natural form showed that their proteolytic and calcium-decreasing activities were at the same level. The successful expression of chymotrypsin C gene paves the way to further mutagenic structure-function studies.
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Authors | Haibo Wang, Duoduo Yuan, Rong Xu, Cheng-Wu Chi |
Journal | Acta biochimica et biophysica Sinica
(Acta Biochim Biophys Sin (Shanghai))
Vol. 43
Issue 7
Pg. 568-75
(Jul 2011)
ISSN: 1745-7270 [Electronic] China |
PMID | 21659382
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- DNA, Complementary
- Recombinant Proteins
- chymotrypsinogen C
- Chymotrypsinogen
- Serine Endopeptidases
- Chymotrypsin
- chymotrypsin C
- Metalloendopeptidases
- peptidyl-Lys metalloendopeptidase
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Topics |
- Amino Acid Sequence
- Animals
- Chymotrypsin
(genetics, isolation & purification)
- Chymotrypsinogen
(chemistry, isolation & purification)
- Cloning, Molecular
- DNA, Complementary
(genetics)
- Escherichia coli
(genetics)
- Inclusion Bodies
(genetics)
- Metalloendopeptidases
(metabolism)
- Molecular Sequence Data
- Pancreas
(enzymology)
- Recombinant Proteins
(chemistry, isolation & purification)
- Serine Endopeptidases
(chemistry, isolation & purification)
- Swine
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