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学科主题: 物理化学
题名: RP-RPLC Multidimensional Separation with High Orthogonality for large-scale Phosphprotemome Analysis
作者: Song CX(宋春霞) ;  Ye ML(叶明亮) ;  Jiang XN(江新宁) ;  Chen R(陈锐) ;  Zou HF(邹汉法)
会议名称: 25th International Symposium on Microscale Bioseparations MSB 2010
会议日期: 2010-3-21
出版日期: 2010-03-21
会议地点: 捷克
通讯作者: 邹汉法
部门归属: 十八室
主办者: 捷克分析化学研究所
摘要: Protein phosphorylation regulates a series of important biological processes in eukaryotes. However, the phosphorylation sites found up to now are far below than that actual existence in proteins due to the extreme complexity of proteome sample. Here a new RP-RPLC 2D separation approach was developed for phosphopeptide separation. By this approach, a large number of fractions were collected from the first dimensional RPLC separation at high pH. And then these fractions were pooled every two fractions with equal time interval, one from early eluted section and the other one from the later eluted section. The pooled fractions were finally submitted to RPLC-MS/MS analysis at low pH. It was found the resulting 2D separation was highly orthogonal and yielded more than 30% phosphopeptide identifications over the conventional RP-RPLC approach. This approach took advantage of the high resolution of RPLC and new fractionation format to improve the orthogonality of two dimensional separations, and provided an highly efficient separation of phosphopeptides for large-scale phosphorylation analysis The developed multidimensional separation approach was integrated to phosphopeptide identification with high-resolution mass spectrometry (LTQ Orbitrap) for large-scale phosphoproteome analysis of human liver, which resulted in the identification of about 10, 000 phosphorylation sites in human liver by controlling FDR less than 1%. Though the experiments are not finished yet, it is already become the largest dataset for human liver phosphoproteome.
语种: 中文
内容类型: 会议论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/114032
Appears in Collections:中国科学院大连化学物理研究所_会议论文

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Recommended Citation:
Song CX,Ye ML,Jiang XN,et al. RP-RPLC Multidimensional Separation with High Orthogonality for large-scale Phosphprotemome Analysis[C]. 见:25th International Symposium on Microscale Bioseparations MSB 2010. 捷克. 2010-3-21.
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