Evaluation of Different Stationary Phases in the Separation of Inter-Cross-Linked Peptides

Journal of Proteome Research
Zixiang FangSaiful M Chowdhury

Abstract

Chemical cross-linking coupled with mass spectrometry (MS) is becoming a routinely and widely used technique for depicting and constructing protein structures and protein interaction networks. One major challenge for cross-linking/MS is the determination of informative low-abundant inter-cross-linked products, generated within a sample of high complexity. A C18 stationary phase is the conventional means for reversed-phase (RP) separation of inter-cross-linked peptides. Various RP stationary phases, which provide different selectivities and retentions, have been developed as alternatives to C18 stationary phases. In this study, two phenyl-based columns, biphenyl and fluorophenyl, were investigated and compared with a C18 phase for separating BS3 (bis(sulfosuccinimidyl)suberate) cross-linked bovine serum albumin (BSA) and myoglobin by bottom-up proteomics. Fractions from the three columns were collected and analyzed in a linear ion trap (LIT) mass spectrometer for improving detection of low abundant inter-cross-linked peptides. Among these three columns, the fluorophenyl column provides additional ion-exchange interaction and exhibits unique retention in separating the cross-linked peptides. The fractioned data was analyzed in pL...Continue Reading

References

May 17, 2000·Proceedings of the National Academy of Sciences of the United States of America·M M YoungG Dollinger
Aug 2, 2003·Journal of the American Society for Mass Spectrometry·Birgit SchillingMalin M Young
Jul 28, 2004·Journal of the American Society for Mass Spectrometry·Bill X HuangChhabil Dass
Oct 1, 2005·Analytical Chemistry·Martin GilarJohn C Gebler
Jun 28, 2007·Nature Reviews. Molecular Cell Biology·Anne-Claude GingrasRuedi Aebersold
Aug 24, 2010·Mass Spectrometry Reviews·Evgeniy V Petrotchenko, Christoph H Borchers
Nov 3, 2010·Journal of Separation Science·Isabel C S F JardimCarol H Collins
Jun 24, 2011·Molecular & Cellular Proteomics : MCP·Chunxiang ZhengJames E Bruce
Sep 1, 2011·Analytical and Bioanalytical Chemistry·Bogusław Buszewski, Sylwia Noga
Nov 1, 2011·Journal of the American Society for Mass Spectrometry·Michael GötzeAndrea Sinz
Jan 31, 2012·Molecular & Cellular Proteomics : MCP·Alexander LeitnerRuedi Aebersold
Feb 7, 2012·Proceedings of the National Academy of Sciences of the United States of America·Keren LaskerWolfgang Baumeister
Feb 14, 2012·Rapid Communications in Mass Spectrometry : RCM·Romy FritzscheAndrea Sinz
Jul 10, 2012·Nature Methods·Bing YangMeng-Qiu Dong
Feb 2, 2015·Journal of Proteomics·Diogo B LimaPaulo C Carvalho
Mar 27, 2015·Journal of Proteome Research·Michael R HoopmannRobert L Moritz
Jul 2, 2015·Proceedings of the National Academy of Sciences of the United States of America·Antje AufderheideFriedrich Förster
Jan 6, 2017·Journal of Proteome Research·Verena TinnefeldRené P Zahedi

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Citations

Feb 8, 2020·Journal of the American Society for Mass Spectrometry·Jayanta K ChakrabartySaiful M Chowdhury
Dec 21, 2021·Journal of the American Society for Mass Spectrometry·Zixiang FangSaiful M Chowdhury

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