Kinetics of acid-mediated disassembly of the b-subunit pentamer of escherichia-coli heat-labile enterotoxin - molecular-basis of ph stability

Ruddock, Lloyd W. and Ruston, Stephen P. and Kelly, Sharon M. and Price, Nicholas C. and Freedman, Robert B. and Hirst, Timothy R. (1995) Kinetics of acid-mediated disassembly of the b-subunit pentamer of escherichia-coli heat-labile enterotoxin - molecular-basis of ph stability. Journal of Biological Chemistry, 270 (50). pp. 29953-29958. ISSN 0021-9258. (The full text of this publication is not available from this repository)

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Abstract

The B-subunit pentamer of Escherichia coli heat-labile enterotoxin (EtxB) is highly stable, maintaining its quaternary structure in a range of conditions that would normally be expected to cause protein denaturation, In this paper the structural stability of EtxB has been studied as a function of pH by electrophoretic, immunochemical, and spectroscopic techniques, Disassembly of the cyclic pentameric structure of human EtxB occurs only below pH 2. As determined by changes in intrinsic fluorescence this process follows first-order kinetics, with the rate constant for disassembly being proportional to the square of the H+ ion concentration, and with an activation energy of 155 kJ mol(-1). A C-terminal deletion mutant, hEtxB214, similarly shows first-order kinetics for disassembly but with a higher pH threshold, resulting in disassembly being seen at pH 3.4 and below. These findings are consistent with the rate-limiting step for disassembly of human EtxB being the simultaneous disruption of two interfaces by protonation of two C-terminal carboxylates. By comparison, disassembly of the B-subunit of cholera toxin (CtxB), a protein which shows 80% sequence identity with EtxB, exhibits a much lower stability to acid conditions; with disassembly of CtxB occurring below pH 3.9, with an activation energy of 81 kJ mol(-1). Reasons for the observed differences in acid stability are discussed, and the implications of these findings to the development of oral vaccines using EtxB and CtxB are considered.

Item Type: Article
Subjects: Q Science
Depositing User: I.T. Ekpo
Date Deposited: 29 Jun 2011 09:27
Last Modified: 28 May 2014 08:31
Resource URI: http://kar.kent.ac.uk/id/eprint/19003 (The current URI for this page, for reference purposes)
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