Determination of Protein Thiol Reduction Potential by Isotope Labeling and Intact Mass Measurement

Analytical Chemistry
Sophie E ThurlowColin J Campbell

Abstract

Oxidation/reduction of thiol residues in proteins is an important type of post-translational modification that is implicated in regulating a range of biological processes. The nature of the modification makes it possible to define a quantifiable electrochemical potential (E(⊕)) for oxidation/reduction that allows cysteine-containing proteins to be ranked based on their propensity to be oxidized. Measuring oxidation of cysteine residues in proteins is difficult using standard electrochemical methods, but top-down mass spectrometry recently has been shown to enable the quantification of E(⊕) for thiol oxidations. In this paper, we demonstrate that mass spectrometry of intact proteins can be used in combination with an isotopic labeling strategy and an automated data analysis algorithm to measure E(⊕) for the thiols in both E. coli Thioredoxin 1 and human Thioredoxin 1. Our methodology relies on accurate mass measurement of proteins using liquid chromatography-mass spectroscopy (LC-MS) analyses and does not necessarily require top-down fragmentation. In addition to analyzing homogeneous protein samples, we also demonstrate that our methodology can be used to determine thiol E(⊕) measurements in samples that contain mixtures of pro...Continue Reading

References

Jan 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·J S StamlerJ Loscalzo
Nov 23, 2000·Nature·T Finkel, N J Holbrook
Jun 21, 2003·The Journal of Biological Chemistry·Walter H WatsonDean P Jones
Dec 5, 2009·ACS Chemical Biology·Candice E Paulsen, Kate S Carroll
Dec 17, 2009·Biochemical and Biophysical Research Communications·Rajib SenguptaDetcho A Stoyanovsky
Jan 12, 2010·The Journal of Biological Chemistry·Simon FourquetMichel B Toledano
Oct 23, 2010·Journal of Internal Medicine·D P Jones
Jul 1, 1996·Analytical Chemistry·A L Rockwood, S L Van Orden
Mar 21, 2012·The Journal of Immunology : Official Journal of the American Association of Immunologists·Ben C KingAnna M Blom
May 23, 2012·Free Radical Biology & Medicine·Venkatesh MallikarjunColin J Campbell
Feb 5, 2013·Analytical Chemistry·David P A KilgourPeter B O'Connor
Aug 14, 2013·Rapid Communications in Mass Spectrometry : RCM·David P A KilgourPeter B O'Connor

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Citations

Feb 6, 2017·Current Protocols in Protein Science·Gregory A Grant
Aug 2, 2019·The Journal of Biological Chemistry·Silvia NapolitanoRudi Glockshuber
Feb 1, 2020·Journal of the American Society for Mass Spectrometry·Kelly J GallagherDavid J Clarke

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