A monoclonal antibody to avidin dissociates quaternary structure and curtails biotin binding to avidin and streptavidin

Subramanian, N and Subramanian, S and Karande, AA and Adiga, PR (1997) A monoclonal antibody to avidin dissociates quaternary structure and curtails biotin binding to avidin and streptavidin. In: Archives of Biochemistry and Biophysics, 344 (2). pp. 281-288.

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Abstract

An anti-avidin mAb, viz., H12G4, is shown to release bound biotin in a dose-dependent manner from holoavidin and holostreptavidin and inhibit the binding of ligand to the two apoproteins. The release of biotin by this mAb is accompanied by quenching of ligand-induced enhanced fluorescence of the FITC-avidin conjugate. In terms of mechanism of release of bound biotin, we demonstrate that on binding to the Fab fragment of the mAb, the native tetrameric holoavidin undergoes dissociation progressively with time to monomers with no bound biotin associated with the latter, Based on the immunoreactivity associated with defined overlapping fragments of avidin obtained by chemical cleavage, the epitope recognized by mAb H12G4 has been localized to residues 58-96 of the primary sequence. By pepscan method of epitope mapping, this mAb is shown to identify a minimal core sequence of (87)RNGK(90); in avidin and a corresponding sequence of (84)RNAN(87) in streptavidin.

Item Type:	Journal Article
Publication:	Archives of Biochemistry and Biophysics
Publisher:	Elsevier Science
Additional Information:	Copyright of this article belongs to Elsevier Science.
Keywords:	Monoclonal antibody;biotin binders;ligand binding;inhibition; epitope mapping
Department/Centre:	Division of Biological Sciences > Biochemistry
Date Deposited:	10 Feb 2010 07:10
Last Modified:	01 Mar 2012 05:42
URI:	http://eprints.iisc.ac.in/id/eprint/18282

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