In order to find candidate proteins that are potentially associated with the phenotype in neonatal rat cardiomyocytes treated with phenylephrine(PE), the differential proteins expression pattern after α-adrenergic receptor stimulation in neonatal cardiomyocytes was analyzed by using a series of methods,including two dimensional polyacrylamide gel electrophoresis,PDQuest 2-DE software analysis and peptide mass fingerprint mapping based on MALDI-TOF-MS.The good two-dimensional pattern with a high resolution and reproducibility was obtained.Totally 14 differential protein spots were found. Out of these,11 proteins were preliminarily identified.Desmin,Txndc5,etc.were found to be up-regulated in PE-treated cardiomyocytes,while caspase-11, 4-hydroxyphenylpyruvic acid dioxygenase,etc.were found to be down-regulated.It suggests that the differential expression analysis of proteome may be useful to the further study of the related proteins and the molecular markers of cardiovascular diseases.
The proteomic maps of human ovarian adenocarcinoma cell line in Human COC1 and its cell subline COC1/DDP were produced by 2-DE SDS-PAGE.The image analysis revealed obviously differential protein expression of the cells in response to cisplatin treatment.Approximate 1 300 protein spots were resolved in the two ranges of pI(3—10).The spots which have 10-fold difference in light density were digested by in-gel(enzyme),identified by matrix assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS) and finally detected by Profound and Mascot search engine in IDI databases.Seven proteins with obviously altered abundances were detected specifically,including four up-regulated protein spots and three down-regulated protein spots.The proteins differentially expressed in the parent cell line COC1 and its subline COC1/DDP were considered to be important for the therapy and pathogenesis.
The protein profiles of the drug-resistant cell line K562/A02 and its sensitive parental cell line K562 were compared by using high resolution two-dimensional gel electrophoresis with IPG strip of pH 4\^7\5\^9. One protein spot was found to be differentially expressed, which was identified as the soluble resistance-related calcium binding protein(sorcin). Tandem mass spectrometric analysis of one tryptic peptide by a TOF-TOF analyzer as well as the western blot analysis substantiates the result derived from peptide mapping fingerprint data.