Detection of phosphopeptides using Fe(III)-nitrilotriacetate complexes immobilized on a MALDI plate

Analytical Chemistry
Jamie D DunnMerlin L Bruening

Abstract

Metal affinity complexes were chemically grafted onto the surface of gold matrix-assisted laser desorption/ionization (MALDI) plates by coupling a derivative of nitrilotriacetate (NTA) to immobilized poly(acrylic acid) (PAA) and subsequently forming the Fe(III)-NTA complex. The immobilized complexes can adsorb phosphorylated peptides preferentially from protein digests; deposition of digests on these surface-modified plates, followed by rinsing with an acetic acid solution, addition of matrix, and subsequent analysis by MALDI MS, resulted in mass spectra dominated by peaks corresponding to phosphopeptides. In the case of analyzing a tryptic digest of beta-casein, conventional MALDI MS revealed only one monophosphopeptide, while use of the Fe(III)-NTA-PAA-modified plate resulted in strong signals due to two additional tetraphosphorylated species. The diminution or elimination of signals due to nonphosphorylated species also greatly simplified the identification of phosphopeptides during analysis of ovalbumin digests and myoglobin digests spiked with an equimolar mixture of angiotensin and phosphoangiotensin. The matrix 2',4',6'-trihydroxyacetophenone mixed with diammonium hydrogen citrate proved to be much better than alpha-cyan...Continue Reading

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Citations

Nov 7, 2006·Analytical and Bioanalytical Chemistry·Chensong PanHanfa Zou
Jul 10, 2009·Journal of the American Society for Mass Spectrometry·Jong-Seo KimHie-Joon Kim
Jul 20, 2010·Annual Review of Analytical Chemistry·Paul W Bohn
Aug 16, 2011·Journal of Proteomics·Ales TichyLenka Hernychova
Apr 10, 2012·Analytica Chimica Acta·Malinda SalimSeetharaman Vaidyanathan
Sep 29, 2011·Trends in Biotechnology·Iris L BatalhaAna C A Roque
Mar 6, 2009·Mass Spectrometry Reviews·Jamie D DunnMerlin L Bruening
Apr 8, 2014·Journal of Proteome Research·Olga ShevchukLothar Jänsch
Feb 12, 2008·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Marcella ChiariClaudio Oldani
Dec 1, 2006·Journal of Mass Spectrometry : JMS
Feb 17, 2019·Journal of Psycholinguistic Research·Isabel R Rodríguez-OrtizDavid Saldaña
Dec 12, 2017·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Se Won BaeWoon-Seok Yeo
Feb 25, 2009·The Analyst·Wei-Han Wang, Merlin L Bruening
Mar 29, 2014·Expert Review of Proteomics·Benjamin Ruprecht, Simone Lemeer
Aug 29, 2009·Journal of Proteome Research·Federico TortaAngela Bachi
Jun 27, 2008·Analytical Chemistry·Jamie D DunnMerlin L Bruening
Jul 23, 2008·Journal of Proteome Research·Michael Schilling, Daniel R Knapp

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