Processing of mutated proinsulin with tetrabasic cleavage sites to bioactive insulin in the non‐endocrine cell line, COS‐7

FEBS Letters

Yanagita, Masahiko²  Nakayama, Kazuhisa¹  Takeuchi, Toshiyuki² 

[1] Gene Experiment Center, University of Tsukuba, Ibaraki 305, Japan;Division of Molecular Endocrinology, Institute for Endocrinology, Gunma University, Maebashi 371, Japan

关键词: Prohormone processing;    Tetrabasic processing site;    Proinsulin;    Insulin;    Furin;    COS-7 cell;   

DOI  :  10.1016/0014-5793(92)81366-T

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The amino acid sequence, Arg⁻⁴-X⁻³-Lys/Arg⁻²-Arg⁻¹ ↓ X⁺¹, is thought to be a consensus processing site for a constitutive secretory pathway in non-endocrine cells. We created a mutant proinsulin DNA with a peptide structure of B chain-Arg-Arg-Lys-Arg-C peptide-Arg-Arg-Lys-Arg-A chain, which compares to the native proinsulin structure of B chain-Arg-Arg-C peptide-Lys-Arg-A chain. When the mutant insulin was expressed in a monkey kidney-derived cell line, COS-7, approximately 60% of the total immunoreactive insulin appeared as mature insulin in the culture medium. This conversion to the mature form was strikingly facilitated by co-expressing the mutant proinsulin with furin, a homologue of the yeast endoprotease, Kex2.

【 授权许可】

Unknown