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Functional expression and characterization of dipeptidyl peptidase IV from the black-bellied hornet Vespa basalis in Sf21 insect cells.
- Source :
-
Bioscience, biotechnology, and biochemistry [Biosci Biotechnol Biochem] 2011; Vol. 75 (12), pp. 2371-5. Date of Electronic Publication: 2011 Dec 07. - Publication Year :
- 2011
-
Abstract
- The maturation of mastoparan B, the major toxin peptide in the venom of Vespa basalis, requires enzymatic cleavage of its prosequence presumably via sequential liberation of dipeptides. The putative processing enzyme, dipeptidyl peptidase IV, was expressed as a glycosylated His-tag fusion protein (rDPP-IV) via the baculovirus expression system. rDPP-IV purified by one-step nickel-affinity chromatography was verified by Western blot and LC-MS/MS analysis. The k(cat)/K(m) of rDPP-IV was determined to be in the range of 10-500 mM(-1)·S(-1) for five synthetic substrates. The optimal temperature and pH for rDPP-IV were determined to be 50 °C and pH 9. Enzymatic activity of rDPP-IV was significantly reduced by 80 and 60% in the presence of sitagliptin and phenylmethylsulfonyl fluoride respectively.
- Subjects :
- Amino Acid Sequence
Animals
Baculoviridae genetics
Cell Line
Chromatography, Affinity
Dipeptidyl Peptidase 4 chemistry
Dipeptidyl Peptidase 4 isolation & purification
Gene Expression
Molecular Sequence Data
Dipeptidyl Peptidase 4 genetics
Dipeptidyl Peptidase 4 metabolism
Wasps enzymology
Wasps genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1347-6947
- Volume :
- 75
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- Bioscience, biotechnology, and biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 22146730
- Full Text :
- https://doi.org/10.1271/bbb.110571