Evaluation of the acid-cleavable isotope-coded affinity tag reagents: application to camptothecin-treated cortical neurons

Journal of Proteome Research
Li-Rong YuTimothy D Veenstra

Abstract

The new generation of isotope-coded affinity tag (ICAT) reagents have been evaluated by labeling an equimolar amount of bovine serum albumin (BSA) with ICAT-12C9 and ICAT-13C9, combining the mixtures, digesting them with trypsin and analyzing the digestate both by muRPLC-tandem MS and by matrix-assisted laser desorption ionization (MALDI) TOF/TOF MS. The use of 13C in place of 2H resulted in both of the labeled peptides having identical elution characteristics in a reversed-phase separation. This similarity in elution allows ICAT-labeled peptides to be effectively analyzed using a muRPLC-MALDI-MS strategy as well. All of the cysteinyl-containing tryptic peptides from BSA were identified with only a 10% variation in the relative abundance measurements between the light and heavy versions of each peptide. A facile method for the removal of contaminants that arise from the cleaved biotin moiety that otherwise interfere with downstream separations and MS analysis has also been developed. The new ICAT reagents were then applied to the analysis of a cortical neuron proteome sample to identify proteins regulated by the antitumor drug, camptothecin.

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Citations

Jul 26, 2007·Chemical Reviews·Tao LiuRichard D Smith
Dec 25, 2010·Journal of Proteome Research·Qing-Rong ChenJaved Khan
Sep 18, 2008·Journal of Translational Medicine·Hua XueMaode Lai
Jun 26, 2009·Expert Opinion on Medical Diagnostics·Jun S WeiJaved Khan
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Oct 14, 2008·Bioorganic Chemistry·Zuly Rivera-MonroyAndrás Guttman
Feb 22, 2008·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Jia TangXiangming Zhang
Oct 9, 2007·Advances in Pediatrics·Stephen W HunsuckerMark W Duncan
Mar 31, 2007·Biochemical and Biophysical Research Communications·Li-Rong YuTimothy D Veenstra
Nov 23, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Mohamed AzarkanGuy Vandenbussche
Feb 14, 2006·Rapid Communications in Mass Spectrometry : RCM·Jeremy E MelansonDevanand M Pinto
Nov 1, 2007·Proteomics. Clinical Applications·Joshua K McBeeRichard S Morrison
Aug 10, 2006·Proteomics·Jeremy E MelansonDevanand M Pinto
Sep 15, 2006·Proteomics·Alexander Leitner, Wolfgang Lindner
Mar 1, 2008·Proteomics. Clinical Applications·Chi Chun WongFeng Chen
Sep 1, 2007·Proteomics. Clinical Applications·Li-Rong YuTimothy D Veenstra
Dec 10, 2013·Journal of Proteomics·Qiang Gu, Li-Rong Yu
Apr 26, 2005·Molecular & Cellular Proteomics : MCP·Robert J Beynon, Julie M Pratt
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Dec 12, 2018·Analytical Chemistry·Albert B Arul, Renã A S Robinson
Sep 23, 2020·Current Protein & Peptide Science·Rocco J Rotello, Timothy D Veenstra

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