【摘要】：等質(zhì)量肽段末端標(biāo)記(Isobaric peptide termini labeling,IPTL)是一種使用輕、重同位素分別對肽段的C端和N端進(jìn)行等重標(biāo)記的技術(shù)。在對使用這種標(biāo)記技術(shù)得到的數(shù)據(jù)進(jìn)行一級(jí)譜分析時(shí),由于肽段的質(zhì)量相同,不會(huì)增加樣本的復(fù)雜性,而在處理二級(jí)譜的數(shù)據(jù)時(shí),可利用成對的b、y離子進(jìn)行分析。本研究利用IPTL方法得到的實(shí)驗(yàn)數(shù)據(jù)設(shè)計(jì)了一種新的打分算法:全部離子打分算法(All ions scoring algorithm,AISA)。AISA在對數(shù)據(jù)進(jìn)行處理時(shí),可以同時(shí)得到定性和定量信息。在Q-Exactive He La和Human-HCC-HL數(shù)據(jù)集上的蛋白定量覆蓋率分別達(dá)到99%和100%。在Q-Exactive He La 2D RPLC數(shù)據(jù)集上,AISA算法鑒定到的PSM、唯一肽段和蛋白質(zhì)分別比Morpheus高15%、26%和22%。在Human-HCC-HL數(shù)據(jù)集上,AISA算法鑒定到的PSM、唯一肽段和蛋白質(zhì)分別比Morpheus高24%、39%和27%。在Q-Exactive He La和Human-HCC-HL數(shù)據(jù)集上蛋白質(zhì)定量比值的平均值非常接近1,分別為1.18和0.90;在0.5~2.0區(qū)間內(nèi)的定量比值分別為91%和94%。
[Abstract]:Isomeric peptide terminal labeling (Isobaric peptide termini labeling,IPTL) is a technique using light and heavy isotopes to label the C-terminal and N-terminal of the peptide respectively. In the first order spectral analysis of the data obtained by using this marking technique, the complexity of the sample will not be increased because of the same quality of the peptide segment, but when the data of the secondary spectrum is processed, the pair of By ions can be used to analyze the data. In this study, a new scoring algorithm is designed based on the experimental data obtained by IPTL method. When the data are processed by (All ions scoring algorithm,AISA). AISA, both qualitative and quantitative information can be obtained. The quantitative protein coverage on Q-Exactive He La and Human-HCC-HL datasets was 99% and 100%, respectively. On the Q-Exactive He La 2D RPLC dataset, the unique peptide and protein of PSM, identified by AISA algorithm were 15% and 22% higher than those of Morpheus, respectively. On the Human-HCC-HL dataset, the unique peptide and protein of PSM, identified by AISA algorithm were 24% and 27% higher than those of Morpheus, respectively. The average values of protein quantitative ratios on Q-Exactive He La and Human-HCC-HL datasets were very close to 1 (1.18 and 0.90, respectively), and the quantitative ratios in the range of 0.5 and 2.0 were 91% and 94%, respectively.
【作者單位】： 北京理工大學(xué)生命學(xué)院;中國科學(xué)院大連化學(xué)物理研究所;
【基金】：國家重大科學(xué)儀器設(shè)備開發(fā)專項(xiàng)基金(No.2012YQ04014005)項(xiàng)目資助~~
【分類號(hào)】：O629.73;O657.63
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本文編號(hào)：2371058