title: Identification of c-type heme-containing peptides using nonactivated immobilized metal affinity chromatography resin enrichment and higher-energy collisional dissociation.
abstract: 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.
authors: Biological Sciences Division and Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, Washington 99352, USA.
authors: Haizhen
authors: H
authors: Zhang
authors: Feng
authors: F
authors: Yang
authors: Wei-Jun
authors: WJ
authors: Qian
authors: Roslyn N
authors: RN
authors: Brown
authors: Yuexi
authors: Y
authors: Wang
authors: Eric D
authors: ED
authors: Merkley
authors: Eric E
authors: EE
authors: Merkley
authors: Jea H
authors: JH
authors: Park
authors: Matthew E
authors: ME
authors: Monroe
authors: Samuel O
authors: SO
authors: Purvine
authors: Ronald J
authors: RJ
authors: Moore
authors: Liang
authors: L
authors: Shi
authors: James K
authors: JK
authors: Fredrickson
authors: Ljiljana
authors: L
authors: Paša-Tolić
authors: Richard D
authors: RD
authors: Smith
authors: Mary S
authors: MS
authors: Lipton
mesh_terms: D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity; D002853:Chromatography, Liquid; D003574:Cytochrome c Group / Q000032:analysis*; D006736:Horses; D008670:Metals / Q000737:chemistry* / Q000378:metabolism; D010446:Peptide Fragments / Q000032:analysis; D040901:Proteomics; D012117:Resins, Synthetic / Q000737:chemistry*; D020592:Shewanella / Q000201:enzymology; D053719:Tandem Mass Spectrometry
chemical_list: D003574:Cytochrome c Group; D008670:Metals; D010446:Peptide Fragments; D012117:Resins, Synthetic