Identification of disulfide-linked peptides by isotope profiles produced by peptic digestion of proteins in 50% (18)O water

Protein Science : a Publication of the Protein Society
Tristan P WallisJeffrey J Gorman

Abstract

Determination of the disulfide-bond arrangement of a protein by characterization of disulfide-linked peptides in proteolytic digests may be complicated by resistance of the protein to specific proteases, disulfide interchange, and/or production of extremely complex mixtures by less specific proteolysis. In this study, mass spectrometry has been used to show that incorporation of (18)O into peptides during peptic digestion of disulfide-linked proteins in 50% (18)O water resulted in isotope patterns and increases in average masses that facilitated identification and characterization of disulfide-linked peptides even in complex mixtures, without the need for reference digests in 100% (16)O water. This is exemplified by analysis of peptic digests of model proteins lysozyme and ribonuclease A (RNaseA) by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) and electrospray ionization (ESI) mass spectrometry (MS). Distinct isotope profiles were evident when two peptide chains were linked by disulfide bonds, provided one of the chains did not contain the C terminus of the protein. This latter class of peptide, and single-chain peptides containing an intrachain disulfide bond, could be identified and characterized by ...Continue Reading

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Citations

Feb 5, 2003·Current Opinion in Biotechnology·Michael B Goshe, Richard D Smith
Dec 9, 2008·Molecular & Cellular Proteomics : MCP·Keyur A DaveJeffrey J Gorman
Jan 5, 2014·Antioxidants & Redox Signaling·Chad R Borges, Nisha D Sherma
Nov 20, 2015·Journal of the American Chemical Society·Victor S LelyveldJack W Szostak
Aug 10, 2018·Expert Review of Proteomics·Per HägglundMichael J Davies
Sep 13, 2005·Rapid Communications in Mass Spectrometry : RCM·Gang Sun, Vernon E Anderson
Oct 4, 2013·Expert Review of Proteomics·Miriam S GoyderFranka Kálmán
Jan 4, 2020·Mass Spectrometry Reviews·Giuseppe Grasso
Feb 26, 2004·The Journal of Biological Chemistry·Tristan P WallisJeffrey J Gorman
Nov 2, 2019·The Journal of Biological Chemistry·Clare L Hawkins, Michael J Davies
Dec 12, 2002·Mass Spectrometry Reviews·Jeffrey J GormanJames J Pitt
Jun 25, 2009·Analytical Chemistry·Sheng Yu HuangYueh Yi Lin
Feb 2, 2002·Science·David LandoMurray L Whitelaw

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