Cloning and expression of human islet amyloid polypeptide in cultured cells

被引：5

作者：

Bhattacharya, Susinjan ^{[1
]}

Latha, J. Naveena Lavanya ^{[1
]}

Kumresan, R. ^{[1
]}

Singh, Shashi ^{[1
]}

机构：

[1] Ctr Cellular & Mol Biol, Hyderabad 500007, Andhra Pradesh, India

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2007年 / 356卷 / 03期

关键词：

recombinant hIAPP; transgenic expression; amyloidogenesis; amyloid fibrils; AFM; thioflavin-T binding; protein purification; apoptosis; amyloid toxicity;

D O I：

10.1016/j.bbrc.2007.03.016

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Efforts to clone amyloidogenic proteins in the cells often have resulted in cell death. We report successful cloning and expression of recombinant human islet amyloid polypeptide (hIAPP) in cultured mammalian cells. Amylin gets secreted, forms fibrils that are toxic to target cells like 0 cells of rat and human. The study involves cloning of full-length amylin in fluorescent protein vector followed by transfection into mammalian cells. The transfected cells with recombinant human amylin, secrete the translated protein corresponding to 37-amino acid native mature IAPP. The mature IAPP secreted out of the cell is purified and characterized by MALDI-TOF/TOF-MS and Western blotting. Purified IAPP forms fibrils as seen by Thioflavin-T fluorescence and AFM, and these fibrils were cytotoxic towards pancreatic cell line RIN5mf cells. (c) 2007 Elsevier Inc. All rights reserved.

引用

页码：622 / 628

页数：7

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