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

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 87RNGK90 in avidin and a corresponding sequence of 84RNAH87 in streptavidin.

journal_name

Arch Biochem Biophys

authors

Subramanian N,Subramanian S,Karande AA,Adiga PR

doi

10.1006/abbi.1997.0196

subject

Has Abstract

pub_date

1997-08-15 00:00:00

pages

281-8

issue

2

eissn

0003-9861

issn

1096-0384

pii

S0003-9861(97)90196-X

journal_volume

344

pub_type

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