MALDI-TOF MS identification of protein of mitochondria from cultured hepatoma cells / 重庆医科大学学报

ABSTRACT

Objective:To explore the optimal conditions associated with proteolytic digestion in 2-DE gel,PMF analysis and database inquiry,and to construct a mass identification strategy in research for mitochondrial comparative proteomics.Methods:The BSA standard protein was incised from 2-DE gel by Coomassie Brilliant Blue dyeing and was mixed with matrix (CHCA),then analyzed by MALDI-TOF MS.The differentially expressed mitochondria protein L9,an unknown protein obtained from hepatoma cells QGY-7703,was identified by condition with BSA optimizing.The PMF was inquired by MS-Fit database.Results:The BSA was identified in Masses Matched number 10/11 and Sequence Overcast 19% of PMF,which proved to the reliability of experimental conditions.By database Inquiry about PMF from L9 protein,OXCT(3-oxoacid CoA transferase 1 precursor)was identified with Masses Matched number 9/13 and Sequence Overcast 24%.The OXCT was in accord with position on gel.Conclusion:This method is applicable to research for mitochondrial comparative proteomics.