High-yield bacterial expression and structural characterization of recombinant human insulin-like growth factor binding protein-2

Swain, Monalisa and Slomiany, Mark G and Rosenzweig, Steven A and Atreya, Hanudatta S (2010) High-yield bacterial expression and structural characterization of recombinant human insulin-like growth factor binding protein-2. In: Archives of Biochemistry and Biophysics, 501 (2). pp. 195-200.

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Abstract

The diverse biological activities of the insulin-like growth factors (IGF-1 and IGF-2) are mediated by the IGF-1 receptor (IGF-1R). These actions are modulated by a family of six IGF-binding proteins (ICFBP-1-6; 22-31 kDa) that via high affinity binding to the IGFs (K-D similar to 300-700 pM) both protect the IGFs in the circulation and attenuate IGF action by blocking their receptor access In recent years, IGFBPs have been implicated in a variety of cancers However, the structural basis of their interaction with IGFs and/or other proteins is not completely understood A critical challenge in the structural characterization of full-length IGFBPs has been the difficulty in expressing these proteins at levels suitable for NMR/X-ray crystallography analysis Here we describe the high-yield expression of full-length recombinant human IGFBP-2 (rhIGFBP-2) in Eschericha coli Using a single step purification protocol, rhIGFBP-2 was obtained with >95% purity and structurally characterized using NMR spectroscopy. The protein was found to exist as a monomer at the high concentrations required for structural studies and to exist in a single conformation exhibiting a unique intra-molecular disulfide-bonding pattern The protein retained full biologic activity. This study represents the first high-yield expression of wild-type recombinant human IGFBP-2 in E coli and first structural characterization of a full-length IGFBP (C) 2010 Elsevier Inc. All rights reserved

Item Type:	Journal Article
Publication:	Archives of Biochemistry and Biophysics
Publisher:	Elsevier Science
Additional Information:	Copyright of this article belongs to Elsevier Science.
Keywords:	Recombinant human IGF-binding protein-2 (rhIGFBP-2); E coli expression; Protein purification; Protein structure; Secondary structure; G-matrix Fourier transform (GFT) NMR
Department/Centre:	Division of Chemical Sciences > NMR Research Centre (Formerly Sophisticated Instruments Facility) Division of Chemical Sciences > Solid State & Structural Chemistry Unit
Date Deposited:	27 Sep 2010 11:28
Last Modified:	27 Sep 2010 11:28
URI:	http://eprints.iisc.ac.in/id/eprint/32486

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