PMID:10497026

Citation	Streaker, ED and Beckett, D (1999) Ligand-linked structural changes in the Escherichia coli biotin repressor: the significance of surface loops for binding and allostery. J. Mol. Biol. 292:619-32
Abstract	The Escherichia coli repressor of biotin biosynthesis (BirA) is an allosteric site-specific DNA-binding protein. BirA catalyzes synthesis of biotinyl-5'-AMP from substrates biotin and ATP and the adenylate serves as the positive allosteric effector in binding of the repressor to the biotin operator sequence. Although a three-dimensional structure of the apo-repressor has been determined by X-ray crystallographic techniques, no structures of any ligand-bound forms of the repressor are yet available. Results of previously published solution studies are consistent with the occurrence of conformational changes in the protein concomitant with ligand binding. In this work the hydroxyl radical footprinting technique has been used to probe changes in reactivity of the peptide backbone of BirA that accompany ligand binding. Results of these studies indicate that binding of biotin to the protein results in protection of regions of the central domain in the vicinity of the active site and the C-terminal domain from chemical cleavage. Biotin-linked changes in reactivity constitute a subset of those linked to adenylate binding. Binding of both bio-5'-AMP and biotin operator DNA suppresses cleavage at additional sites in the amino and carboxy-terminal domains of the protein. Varying degrees of protection of the five surface loops on BirA from hydroxyl radical-mediated cleavage are observed in all complexes. These results implicate the C-terminal domain of BirA, for which no function has previously been known, in small ligand and site-specific DNA binding and highlight the significance of surface loops, some of which are disordered in the apoBirA structure, for ligand binding and transmission of allosteric information in the protein.
Links	PubMed Online version:10.1006/jmbi.1999.3086
Keywords	Adenosine Monophosphate/analogs & derivatives; Adenosine Monophosphate/biosynthesis; Allosteric Regulation; Bacterial Proteins/chemistry; Binding Sites; Biotin/analogs & derivatives; Biotin/biosynthesis; Biotin/metabolism; Carbon-Nitrogen Ligases/chemistry; Cyclic AMP-Dependent Protein Kinases/genetics; DNA/chemistry; DNA-Binding Proteins/chemistry; Escherichia coli/genetics; Escherichia coli Proteins; Gene Expression Regulation, Bacterial; Hydroxyl Radical/metabolism; Ligands; Models, Molecular; Mutation; Protein Binding; Recombinant Fusion Proteins; Repressor Proteins/chemistry; Transcription Factors
edit table

Significance

You can help EcoliWiki by summarizing why this paper is useful

Useful Materials and Methods

You can help Ecoliwiki by describing the useful materials (strains, plasmids, antibodies, etc) described in this paper.

Annotations

<annotationlinks/>

EcoliWiki Links

Add links to pages that link here (e.g. gene, product, method pages)

References

See Help:References for how to manage references in EcoliWiki.