期刊
PROTEOMICS
卷 8, 期 8, 页码 1564-1575出版社
WILEY
DOI: 10.1002/pmic.200700851
关键词
cardiac mitochondria; mass spectrometry; sample preparation; target validation
资金
- NCRR NIH HHS [RR-022371-01, S10 RR022371] Funding Source: Medline
- NHLBI NIH HHS [R01 HL101228, R37 HL063901, R21 HL076526, HL-78109, F32 HL078109, HL-76526, HL-63901, HL-65431, R01 HL063901-09, R01 HL065431-09, HL-80111, P01 HL080111, R01 HL063901, R01 HL065431, P01 HL080111-04, F32 HL078109-01, R21 HL076526-02] Funding Source: Medline
Mitochondria play essential roles in cardiac pathophysiology and the murine model has been extensively used to investigate cardiovascular diseases. In the present study, we characterized murine cardiac mitochondria using an LC/MS/MS approach. We extracted and purified cardiac mitochondria; validated their functionality to ensure the final preparation contains necessary components to sustain their normal function; and subjected these validated organelles to LC/ MS/MS-based protein identification. A total of 940 distinct proteins were identified from murine cardiac mitochondria, among which, 480 proteins were not previously identified by major proteomic profiling studies. The 940 proteins consist of functional dusters known to support oxidative phosphorylation, metabolism, and biogenesis. In addition, there are several other clusters, including proteolysis, protein folding, and reduction/oxidation signaling, which ostensibly represent previously under-appreciated tasks of cardiac mitochondria. Moreover, many identified proteins were found to occupy other subcellular locations, including cytoplasm, ER, and golgi, in addition to their presence in the mitochondria. These results provide a comprehensive picture of the murine cardiac mitochondrial proteome and underscore tissue- and species-specification. Moreover, the use of functionally intact mitochondria insures that the proteomic observations in this organelle are relevant to its normal biology and facilitates decoding the interplay between mitochondria and other organelles.
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