1. Identification of c-Type Heme-Containing Peptides Using Nonactivated Immobilized Metal Affinity Chromatography Resin Enrichment and Higher-Energy Collisional Dissociation.
- Author
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Zhang, Haizhen, Feng Yang, Wei-Jun Qian, Brown, Roslyn N., Yuexi Wang, Merkley, Eric E., Park, Jea H., Monroe, Matthew E., Purvine, Samuel O., Moore, Ronald J., Liang Shi, Fredrickson, James K., Paša-Tolić, Ljiljana, Smith, Richard D., and Lipton, Mary S.
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CYTOCHROMES , *CHARGE exchange , *APOPTOSIS , *PROKARYOTES , *EUKARYOTIC cells , *PEPTIDE fractionation - 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. [ABSTRACT FROM AUTHOR]
- Published
- 2011
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