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A novel approach for over-expression, characterization, and isotopic enrichment of a homogeneous species of acyl carrier protein from Plasmodium falciparum

Sharma, Shailendra Kumar and Modak, Rahul and Sharma, Shilpi and Sharma, Alok Kumar and Sarma, Siddhartha P and Surolia, Avadhesha and Surolia, Namita (2005) A novel approach for over-expression, characterization, and isotopic enrichment of a homogeneous species of acyl carrier protein from Plasmodium falciparum. In: Biochemical and Biophysical Research Communications, 330 (4). pp. 1019-1026.

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Abstract

Acyl carrier protein (ACP) plays a central role in fatty acid biosynthesis by transferring the acyl groups from one enzyme to another for the completion of the fatty acid synthesis cycle. Holo-ACP is the obligatory substrate for the synthesis of acyl-ACPs which act as the carrier and donor for various metabolic reactions. Despite its interactions with numerous proteins in the cell, its mode of interaction is poorly understood. Here, we report the over-expression of PfACP in minimal medium solely in its holo form and in high yield.Expression in minimal media provides a means to isotopically label PfACP for high resolution multi-nuclear and multi-dimensional NMR studies. Indeed, the proton-nitrogen correlated NMR spectrum exhibits very high chemical shift dispersion and resolution. We also show that holo-PfACP thus expressed is amenable to acylation reactions using Escherichia coli acyl-ACP synthetase as well as by standard chemical methods.

Item Type: Journal Article
Additional Information: Copyright for this article belongs to Elsevier.
Keywords: Protein expression;Plasmodium falciparum;Apo-ACP;Holo-ACP;Acyl-ACPs;NMR
Department/Centre: Division of Biological Sciences > Molecular Biophysics Unit
Date Deposited: 08 Jun 2005
Last Modified: 19 Sep 2010 04:19
URI: http://eprints.iisc.ernet.in/id/eprint/3291

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