Identification and sequencing analysis of intact proteins via collision-induced dissociation and quadrupole time-of-flight mass spectrometry

Journal of Mass Spectrometry : JMS
Jennifer F Nemeth-Cawley, Jason C Rouse

Abstract

Identifying unknown proteins has become a central focal point for proteomic and biopharmaceutical development laboratories. Our laboratory investigated using quadrupole time-of-flight mass spectrometry (Qq/TOFMS) for the analysis of intact proteins for the purpose of identifying unknowns while limiting the number of sample-handling steps between protein extraction and identification. Eight standard proteins, both unmodified and disulfide-bonded and ranging in mass from 5 to 66 kDa, were analyzed using nanoelectrospray and collision-induced dissociation to generate peptide sequence tags. An MS analysis, followed by MS/MS analyses on two to five individual protein charge states, were obtained to make an identification. Peptide sequence tags were extracted from the MS/MS data and used, in conjunction with molecular mass and source origin, to obtain protein identifications using the web-based search engine ProteinInfo (www.proteometrics.com). All of the proteins were unambiguously identified from the input data, after which, all of the major product ions were identified for structural information. In most cases, N- and/or C-terminal ions, and also stretches of consecutive product ions from the protein interior, were observed. This ...Continue Reading

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Citations

Jul 19, 2002·Journal of Mass Spectrometry : JMS·Gavin E Reid, Scott A McLuckey
Jul 22, 2008·Glycoconjugate Journal·François FenailleNicolas Bihoreau
Jul 19, 2005·Journal of the American Society for Mass Spectrometry·Nadezhda A GalevaThomas C Squier
Nov 8, 2005·Journal of the American Society for Mass Spectrometry·Judith A JebanathirajahPeter B O'Connor
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Jun 5, 2003·Journal of the American Society for Mass Spectrometry·Mario ThevisJoseph A Loo
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Mar 20, 2003·Journal of Mass Spectrometry : JMS·Jason M Hogan, Scott A McLuckey
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Nov 26, 2005·Rapid Communications in Mass Spectrometry : RCM·Rupasri Mandal, Xing-Fang Li
Nov 23, 2006·Rapid Communications in Mass Spectrometry : RCM·Putchen Dakshinamoorthy Deepalakshmi
Dec 15, 2005·Rapid Communications in Mass Spectrometry : RCM·Peter B O'connorCheng Zhao
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Oct 31, 2018·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Delphine VincentSimone Rochfort
Mar 27, 2010·Analytical Chemistry·Paul DrogarisPierre Thibault

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