Production of recombinant salmon calcitonin by amidation of precursor peptide using enzymatic transacylation and photolysis in vitro | Health & Environmental Research Online (HERO)

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Citation
Tags

HERO ID

5195054

Reference Type

Journal Article

Title

Production of recombinant salmon calcitonin by amidation of precursor peptide using enzymatic transacylation and photolysis in vitro

Author(s)

Hong, D; Mingqiang, Z; Min, L; Changqing, C; Jifang, M

Year

2000

Is Peer Reviewed?

Yes

Journal

Biochemical and Biophysical Research Communications
ISSN: 0006-291X
EISSN: 1090-2104

Volume

267

Issue

Page Numbers

362-367

Language

English

PMID

10623625

DOI

10.1006/bbrc.1999.1961

Web of Science Id

WOS:000084990000062

Abstract

The C terminal amidation is required for full biological activity of salmon calcitonin (sCT). We constructed BL21(DE3)/pGEX-sCT-Ala, an engineering Escherichia coli strain. The soluble fusion protein of GST-sCT-Ala expressed from BL21(DE3)/pGEX-sCT-Ala was purified by affinity chromatography after high density, high expression culture and sonication of bacteria. Following S-sulfonation of the fusion protein, the 33 alanine-extended peptides were released from the fusion protein by cyanogen bromide. The S-sulfonated precursor peptide was transacylated by CPD-Y, o-PNGA as a nucleophile, to produce photosensitive SO(-)(3)-sCT-o-PNGA. After photolysis and folding, the biological activity of sCT was assayed as standard.