Identification of c-type heme-containing peptides using nonactivated immobilized metal affinity chromatography resin enrichment and higher-energy collisional dissociation.
Zhang, Haizhen; Yang, Feng; Qian, Wei-Jun; Brown, Roslyn N; Wang, Yuexi; Merkley, Eric D; Merkley, Eric E; Park, Jea H; Monroe, Matthew E; Purvine, Samuel O; Moore, Ronald J; Shi, Liang; Fredrickson, James K; Pasa-Tolic, Ljiljana; Smith, Richard D; Lipton, Mary S.
Anal Chem;
83(19): 7260-8, 2011 Oct 01.
Artigo
em Inglês
| MEDLINE | ID: mdl-21740036
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.
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