Document Detail


Cloning and expression of human islet amyloid polypeptide in cultured cells.
MedLine Citation:
PMID:  17374526     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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 beta 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.
Authors:
Susinjan Bhattacharya; J Naveena Lavanya Latha; R Kumresan; Shashi Singh
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2007-03-12
Journal Detail:
Title:  Biochemical and biophysical research communications     Volume:  356     ISSN:  0006-291X     ISO Abbreviation:  Biochem. Biophys. Res. Commun.     Publication Date:  2007 May 
Date Detail:
Created Date:  2007-04-05     Completed Date:  2007-06-06     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0372516     Medline TA:  Biochem Biophys Res Commun     Country:  United States    
Other Details:
Languages:  eng     Pagination:  622-8     Citation Subset:  IM    
Affiliation:
Centre for Cellular and Molecular Biology, Hyderabad 500007, India.
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MeSH Terms
Descriptor/Qualifier:
Amyloid / biosynthesis,  genetics*
Animals
CHO Cells
COS Cells
Cell Survival
Cells, Cultured
Cercopithecus aethiops
Cloning, Molecular
Cricetinae
Cricetulus
Escherichia coli / metabolism
Humans
Microscopy, Atomic Force
Rats
Spectrometry, Fluorescence
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Thiazoles
Transfection
Chemical
Reg. No./Substance:
0/Amyloid; 0/Thiazoles; 106602-62-4/amylin; 2390-54-7/thioflavin T

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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