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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, L. M. (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. doi:10.1529/biophysj.105.076455
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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: |
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| Volume: | Vol.90 | ||||
| Number: | No.9 | ||||
| Page Range: | pp. 3246-3254 | ||||
| DOI: | 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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