Functional expression of horsegram (Dolichos biflorus) Bowman–Birk inhibitor and its self-association.

Deepa, G. Muricken (2010) Functional expression of horsegram (Dolichos biflorus) Bowman–Birk inhibitor and its self-association. Biochimica et Biophysica Acta, 1804. pp. 1413-1423. ISSN 0006-3002

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Abstract

Horsegram (Dolichos biflorus), a protein-rich leguminous pulse, native to Southeast Asia and tropical Africa, contains multiple forms of Bowman–Birk inhibitors. The major Bowman–Birk inhibitor from horsegram (HGI-III) was cloned and functionally expressed in Escherichia coli (rHGI), which moved as a dimer in solution similar to the natural inhibitor. The biochemical characterization of rHGI also points to its close resemblance with HGI-III not only in its structure but also in its inhibitory characteristics. To explore the electrostatic interactions involved in the dimerization, a site-directed mutagenesis approach was used. The role of reactive site residue K24 and the C-terminal Asp in the structure and stability of the dimer was accomplished by mutating K24 and D75/76. The mutants produced in this study confirm that the selfassociation of HGI-III is indeed due to the electrostatic interaction between K24 of one monomer and D75/76 of the second monomer, in agreement with our previous data. The functional expression of a Bowman–Birk inhibitor minus a fusion tag serves as a platform to study the structural and functional effects of the special pattern of seven conserved disulphide bridges.

Item Type:	Article
Uncontrolled Keywords:	Recombinant horsegram inhibitor, Cloning Site-directed mutagenesis, Monomer–dimer interaction C-terminal Asp
Subjects:	500 Natural Sciences and Mathematics > 04 Chemistry and Allied Sciences > 16 Enzyme Chemistry 600 Technology > 08 Food technology > 22 Legumes-Pulses
Divisions:	Protein Chemistry and Technology
Depositing User:	Food Sci. & Technol. Information Services
Date Deposited:	13 Nov 2013 04:59
Last Modified:	13 Nov 2013 04:59
URI:	http://ir.cftri.res.in/id/eprint/11282

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