Comparison of metal and metal oxide media for phosphopeptide enrichment prior to mass spectrometric analyses.

Journal of the American Society for Mass Spectrometry
Matthew B GatesLeesa J Deterding

Abstract

Several affinity resins consisting of ionic metals or metal oxides were investigated for their phosphopeptide enrichment capabilities with subsequent mass spectrometric analyses. Commercially-available enrichment metal oxide affinity chromatography (MOAC) resins using manufacturer's and/or published protocols were compared and evaluated for the most efficient and selective method that could be implemented as a standard enrichment procedure. From these comparative analyses, using a tryptic digest of casein proteins, it was determined that in our hands, two of the resins out-performed the others based on a variety of criteria, including the number of phosphorylation sites identified during MS analyses, the lower numbers of nonspecifically bound peptides observed, and the limits of detection. Applicability of these enrichment resins to a complex biological mixture was investigated. For this work, a mixture of avian histones was digested, subjected to titanium dioxide phosphopeptide enrichment, and analyzed by mass spectrometry. Eight phosphorylated tryptic peptides were observed following enrichment and subsequent LC/MS/MS analyses. Of note, seven of the eight phosphopeptides were not observed without titanium dioxide enrichment. ...Continue Reading

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Citations

Oct 18, 2011·Amino Acids·Jan Fíla, David Honys
Jul 24, 2012·Amino Acids·Luisa Beltran, Pedro R Cutillas
Dec 4, 2013·Journal of the American Society for Mass Spectrometry·Francesco LanucaraClaire E Eyers
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Jul 3, 2015·Frontiers in Plant Science·Jinna LiHaiying Li
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Jun 2, 2018·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Guo ZhongPeter W Stacpoole
Mar 28, 2015·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Xuan HuangShuo Wang
Nov 3, 2011·Analytical Chemistry·Carol L Nilsson

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