Intermolecular interactions between the a-subunit and b-subunit of heat-labile enterotoxin from escherichia-coli promote holotoxin assembly and stability invivo

Streatfield, S.J. and Sandkvist, M. and Sixma, T.K. and Bagdasarian, M. and Hol, W.G.J. and Hirst, Timothy R. (1992) Intermolecular interactions between the a-subunit and b-subunit of heat-labile enterotoxin from escherichia-coli promote holotoxin assembly and stability invivo. Proceedings of the National Academy of Sciences of the United States of America, 89 (24). pp. 12140-12144. ISSN 0027-8424. (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided)

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Official URL
http://dx.doi.org/10.1073/pnas.89.24.12140

Abstract

Cholera toxin and the related heat-labile enterotoxin (LT) produced by Escherichia coli consist of a holotoxin of one A subunit and five B subunits (AB5). Here we investigate the domains of the A subunit (EtxA) of E. coli LT which influence the events of B-subunit (EtxB) oligomerization and the formation of a stable AB5 holotoxin complex. We show that the C-terminal 14 amino acids of the A subunit comprise two functional domains that differentially affect oligomerization and holotoxin stability. Deletion of the last 14 amino acids (-14) from the A subunit resulted in a molecule that was significantly impaired in its capacity to promote the assembly of a mutant B subunit, EtxB191.5. In contrast, deletion of the last four amino acids (-4) from the A subunit gave a molecule that retained such a capacity. This suggests that C-terminal residues within the -14 to -4 region of the A subunit are important for promoting the oligomerization of EtxB. In addition, we demonstrate that the truncated A subunit lacking the last 4 amino acids was unable to form a stable AB5 holotoxin complex even though it promoted B-subunit oligomerization. This suggests that the last 4 residues of the A subunit function as an "anchoring" sequence responsible for maintaining the stability of A/B subunit interaction during holotoxin assembly. These data represent an important example of how intermolecular interactions between polypeptides in vivo can modulate the folding and assembly of a macromolecular complex.

Item Type: Article
Uncontrolled keywords: protein export; protein folding; endoplasmic reticulum retention signal; cholera toxin
Subjects: Q Science
Divisions: Faculties > Science Technology and Medical Studies > School of Biosciences
Depositing User: O.O. Odanye
Date Deposited: 31 Aug 2009 21:47
Last Modified: 19 Jun 2014 15:23
Resource URI: https://kar.kent.ac.uk/id/eprint/22247 (The current URI for this page, for reference purposes)
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