Top-Down Proteomic Identification of Furin-Cleaved α-Subunit of Shiga Toxin 2 from Escherichia coli O157 : H7 Using MALDI-TOF-TOF-MSMS
Joint Authors
Sultan, Omar
Fagerquist, Clifton K.
Source
Journal of Biomedicine and Biotechnology
Issue
Vol. 2010, Issue 2010 (31 Dec. 2010), pp.1-11, 11 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2011-02-10
Country of Publication
Egypt
No. of Pages
11
Main Subjects
Abstract EN
A method has been developed to identify the α-subunit of Shiga toxin 2 (α-Stx2) from Escherichia coli O157:H7 using matrix-assisted laser desorption/ionization time-of-flight-time-of-flight tandem mass spectrometry (MALDI-TOF-TOF-MS/MS) and top-down proteomics using web-based software developed in-house.
Expression of Stx2 was induced by culturing E.
coli O157:H7 on solid agar supplemented with an antibiotic that elicits the bacterial SOS-response.
Bacterial cell lysates were incubated in the presence of furin, a human enzyme, that cleaves α-Stx2 into A1 (~28 kDa) and A2 (~5 kDa) protein fragments.
A subsequent disulfide reduction step unlinked A1 from A2.
MALDI-TOF-MS of the furin-digested/disulfide-reduced sample showed a peak at mass-to-charge (m/z) 5286 that corresponded to the A2 fragment.
No peak was observed that corresponded to the A1 fragment although its presence was confirmed by bottom-up proteomics.
The peak at m/z 5286 was definitively identified by MALDI-TOF-TOF-MS/MS and top-down proteomics as the A2 fragment of α-Stx2.
American Psychological Association (APA)
Fagerquist, Clifton K.& Sultan, Omar. 2011. Top-Down Proteomic Identification of Furin-Cleaved α-Subunit of Shiga Toxin 2 from Escherichia coli O157 : H7 Using MALDI-TOF-TOF-MSMS. Journal of Biomedicine and Biotechnology،Vol. 2010, no. 2010, pp.1-11.
https://search.emarefa.net/detail/BIM-447409
Modern Language Association (MLA)
Fagerquist, Clifton K.& Sultan, Omar. Top-Down Proteomic Identification of Furin-Cleaved α-Subunit of Shiga Toxin 2 from Escherichia coli O157 : H7 Using MALDI-TOF-TOF-MSMS. Journal of Biomedicine and Biotechnology No. 2010 (2010), pp.1-11.
https://search.emarefa.net/detail/BIM-447409
American Medical Association (AMA)
Fagerquist, Clifton K.& Sultan, Omar. Top-Down Proteomic Identification of Furin-Cleaved α-Subunit of Shiga Toxin 2 from Escherichia coli O157 : H7 Using MALDI-TOF-TOF-MSMS. Journal of Biomedicine and Biotechnology. 2011. Vol. 2010, no. 2010, pp.1-11.
https://search.emarefa.net/detail/BIM-447409
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-447409