【摘要】：為了優(yōu)化肽質(zhì)量指紋譜(peptide mass fingerprint,PMF)鑒定蛋白質(zhì)的生物信息學(xué)分析條件。將牛碳酸酐酶2(carbonic anhydrase-2,CAH2)和人熱休克蛋白70s(Hsp70s)進(jìn)行2-DE分離、酶解,肽段經(jīng)過MALDI-TOFMS分析得到PMF數(shù)據(jù)。選擇Swissprot、MSDB、NCBInr、Random等數(shù)據(jù)庫和MASCOT與MS-Fit搜索引擎,以牛CAH2為模型優(yōu)化搜索參數(shù),結(jié)果表明:Swissprot是適合做蛋白PMF分析的數(shù)據(jù)庫;主要參數(shù)最佳設(shè)置為:漏切位點(diǎn)數(shù)為1個(gè),肽質(zhì)量容錯(cuò)數(shù)為±1Da,同時(shí)肽質(zhì)量類型選擇平均分子質(zhì)量比單同位素質(zhì)量更便于候選蛋白的篩選。最后用人Hsp70s蛋白的PMF數(shù)據(jù)檢驗(yàn)優(yōu)化條件,結(jié)果表明,所選擇的數(shù)據(jù)庫及參數(shù)是可靠的。
[Abstract]:In order to optimize the bioinformatics analysis conditions for protein identification by peptide mass fingerprint (peptide mass fingerprint,PMF). Bovine carbonic anhydrase 2 (carbonic anhydrase-2,CAH2) and human heat shock protein 70 s (Hsp70s) were separated by 2-DE and hydrolyzed. The PMF data were obtained by MALDI-TOFMS analysis. Swissprot,MSDB,NCBInr,Random database and MASCOT and MS-Fit search engine were selected to optimize the search parameters using bovine CAH2 as the model. The results showed that Swissprot was a suitable database for protein PMF analysis. The main parameters are as follows: the number of missing site is 1, the fault-tolerant number of peptide mass is 鹵1Da. meanwhile, the selection of average molecular weight of peptide mass is more convenient than that of single isotope mass to screen candidate protein. Finally, the optimized conditions were tested with the PMF data of human Hsp70s protein. The results showed that the selected database and parameters were reliable.
【作者單位】： 重慶醫(yī)科大學(xué)藥學(xué)院藥物分析教研室 重慶醫(yī)科大學(xué)藥學(xué)院生物化學(xué)與分子生物學(xué)教研室 重慶醫(yī)科大學(xué)藥學(xué)院藥物分析教研室 重慶醫(yī)科大學(xué)藥學(xué)院藥物分析教研室
【基金】：國家自然科學(xué)基金(No.30476021) 教育部博士點(diǎn)(No.教特發(fā)中心函【2004】166號(hào))資助項(xiàng)目
【分類號(hào)】：Q51
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本文編號(hào)：2402813