Metal affinity engineering of proinsulin carrying genetically attached (His)10-X-met affinity tail and removal of the tag by cyanogen bromide

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Title
Metal affinity engineering of proinsulin carrying genetically attached (His)10-X-met affinity tail and removal of the tag by cyanogen bromide
Author(s)
Jung Hun Koh; Wook Joon Chung; Suk Hoon Koh; Byoung Chul Park; Suk Tae Kwon; Chul Ho Kim; Dae Sil Lee
Bibliographic Citation
Bioscience Biotechnology and Biochemistry, vol. 58, no. 9, pp. 1694-1699
Publication Year
1994
Abstract
An E. coli expression clone coding for human proinsulin, which was fused to NH2-terminal beta-galactosidase, was engineered for the separation from host proteins by introducing peptide devices, and for the sequential removal of the fused polypeptide by cyanogen bromide in front of the NH2 terminal residue (methionine) of the human proinsulin gene. Short synthetic genes encoding oligopeptide residues including (Glu)n, (His)n, (Trp)n, and (Ser)n (n = 10 or 11), which have certain characteristic physical properties such as metal-affinity, polarity, hydrophobicity, and hydrophilicity, respectively, were inserted at the junction region of the gene fusion. Interestingly, it was found that among the oligopeptides, the oligohistidine residue as an affinity-tag has greatly facilitated the procedures for FPI purification, particularly in the manner of selective metal-affinity precipitation. The chelating peptide covering the NH2-terminal beta-galactosidase portion could then be removed simply after purification to generate a protein with the natural amino acid sequence of proinsulin by cyanogen bromide.
ISSN
0916-8451
Publisher
T&F (Taylor & Francis)
Type
Article
Appears in Collections:
Synthetic Biology and Bioengineering Research Institute > Genome Editing Research Center > 1. Journal Articles
Critical Diseases Diagnostics Convergence Research Center > 1. Journal Articles
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