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Gas phase characterization of the noncovalent quaternary structure of Cholera toxin and the Cholera toxin B subunit pentamer

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Williams, Jonathan P., Smith, Daniel C. , Green, Brian N., Marsden, Brian D., Jennings, Keith R., Roberts, Lynne M. and Scrivens, James H.. (2006) Gas phase characterization of the noncovalent quaternary structure of Cholera toxin and the Cholera toxin B subunit pentamer. Biophysical Journal, Vol.90 (No.9). pp. 3246-3254. ISSN 0006-3495

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Official URL: http://dx.doi.org/10.1529/biophysj.105.076455

Abstract

Cholera toxin (CTx) is an AB5 cytotonic protein that has medical relevance in cholera and as a novel mucosal adjuvant. Here, we report an analysis of the noncovalent homopentameric complex of CTx B chain (CTx B5) using electrospray ionization triple quadrupole mass spectrometry and tandem mass spectrometry and the analysis of the noncovalent hexameric holotoxin usingelectrospray ionization time-of-flight mass spectrometry over a range of pH values that correlate with those encountered by this toxin after cellular uptake. We show that noncovalent interactions within the toxin assemblies were maintained under both acidic and neutral conditions in the gas phase. However, unlike the related Escherichia coli Shiga-like toxin B5 pentamer (SLTx B), the CTx B5 pentamer was stable at low pH, indicating that additional interactions must be present within the latter. Structural comparison of the CTx B monomer interface reveals an additional α-helix that is absent in the SLTx B monomer. In silico energy calculations support interactions between this helix and the adjacent monomer. These data provide insight into the apparent stabilization of CTx B relative to SLTx B.

Item Type:

Journal Article

Subjects:

Q Science > QR Microbiology > QR180 Immunology

Divisions:

Faculty of Science > Life Sciences (2010- ) > Biological Sciences ( -2010)

Library of Congress Subject Headings (LCSH):

Vibrio cholerae, Cholera toxin, Electrospray ionization mass spectrometry, Hydrogen-ion concentration -- Physiological effect

Journal or Publication Title:

Biophysical Journal

Publisher:

Biophysical Society

ISSN:

0006-3495

Official Date:

1 May 2006

Dates:

Date	Event
1 May 2006	Published

Volume:

Vol.90

Number:

No.9

Page Range:

pp. 3246-3254

Identification Number:

10.1529/biophysj.105.076455

Status:

Peer Reviewed

Access rights to Published version:

Open Access

Funder:

Wellcome Trust (London, England)

Grant number:

063058/Z/00/Z (Wellcome)

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