Document Detail


Heterologous expression and purification of the soybean 7S globulin α' subunit extension region: In vitro evidence of its involvement in cell cholesterol homeostasis.
MedLine Citation:
PMID:  21821129     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
In a previous paper, the biological activity of a 216-amino acid recombinant truncated form of the soybean 7S globulin α' subunit, known to control cholesterol and triglyceride homeostasis, was described. In this work, a shorter version of the polypeptide chain, spanning 142 amino acid residues from the N-terminus and thus exclusively including the so-called extension region, was cloned and overexpressed in Pichia pastoris. The yield of the recombinant polypeptide, which was termed α'E, was 8-fold greater than the previous truncated version. The α'E polypeptide was purified by simple conventional biochemical techniques to make it available for biological assays. Human hepatoma cell lines (Hep G2) were used to monitor the uptake and degradation of labeled low-density lipoproteins (LDL), according to an established procedure. The LDL uptake (+86%) and degradation (+94%) by cells tested at the highest α'E dose (2μM) were similar to those found in cells incubated with 1μM simvastatin, a potent inhibitor of cholesterol biosynthesis. Additionally, the cell response to α'E was found to be dose-dependent. The present findings strongly suggest that this recombinant polypeptide, or a fragment thereof, is the molecular determinant for cholesterol homeostasis and open new prospects for understanding the mechanism involved in this biological response, as a gateway to its utilization in lipid-lowering therapies.
Authors:
Alessandro Consonni; Maria Rosa Lovati; Anna Parolari; Cristina Manzoni; Paolo Morazzoni; Chiara Magni; Marcello Duranti
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-7-29
Journal Detail:
Title:  Protein expression and purification     Volume:  -     ISSN:  1096-0279     ISO Abbreviation:  -     Publication Date:  2011 Jul 
Date Detail:
Created Date:  2011-8-8     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9101496     Medline TA:  Protein Expr Purif     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2011. Published by Elsevier Inc.
Affiliation:
Department of AgriFood Molecular Sciences, Università degli Studi di Milano, Italy.
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