Identification of c-Type Heme-Containing Peptides Using Nonactivated Immobilized Metal Affinity Chromatography Resin Enrichment and Higher-Energy Collisional Dissociation

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• 作者：Haizhen Zhang ; Feng Yang ; Wei-Jun Qian ; Roslyn N. Brown ; Yuexi Wang ; Eric D. Merkley ; Jea H. Park ; Matthew E. Monroe ; Samuel O. Purvine ; Ronald J. Moore ; Liang Shi ; James K. Fredrickson ; Ljiljana Pa拧a-Toli膰 ; Richard D. Smith ; Mary S. Lipton
• 刊名：Analytical Chemistry
• 出版年：2011
• 出版时间：October 1, 2011
• 年：2011
• 卷：83
• 期：19
• 页码：7260-7268
• 全文大小：650K
• 年卷期：v.83,no.19(October 1, 2011)
• ISSN：1520-6882

文摘

The c-type cytochromes play essential roles in many biological activities of both prokaryotic and eukaryotic cells, including electron transfer, enzyme catalysis, and induction of apoptosis. We report a novel enrichment strategy for identifying c-type heme-containing peptides that uses nonactivated IMAC resin. The strategy demonstrated at least 7-fold enrichment for heme-containing peptides digested from a cytochrome c protein standard, and quantitative linear performance was also assessed for heme-containing peptide enrichment. Heme-containing peptides extracted from the periplasmic fraction of Shewanella oneidensis MR-1 were further identified using higher-energy collisional dissociation tandem mass spectrometry. The results demonstrated the applicability of this enrichment strategy to identify c-type heme-containing peptides from a highly complex biological sample and, at the same time, confirmed the periplasmic localization of heme-containing proteins during suboxic respiration activities of S. oneidensis MR-1.