Protein phosphorylation and expression profiling by Yin-Yang multidimensional liquid chromatography (Yin-Yang MDLC) mass spectrometry

	Protein phosphorylation and expression profiling by Yin-Yang multidimensional liquid chromatography (Yin-Yang MDLC) mass spectrometry
	Dai, J; Jin, WH; Sheng, QH; Shieh, CH; Wu, JR; Zeng, R
刊名	JOURNAL OF PROTEOME RESEARCH
	2007
卷号	6 期号:1 页码:250-262
关键词	protein phosphorylation protein expression strong-cation exchange strong anion exchange Yin-Yang multidimensional liquid chromatography pH elution mass spectrometry
通讯作者	Zeng, R (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Res Ctr Proteome Anal, Inst Biochem & Cell Biol, Shanghai 200031, Peoples R China.,zr@sibs.ac.cn
英文摘要	A system which consisted of multidimensional liquid chromatography (Yin-yang MDLC) coupled with mass spectrometry was used for the identification of peptides and phosphopeptides. The multidimensional liquid chromatography combines the strong-cation exchange (SCX), strong-anion exchange (SAX), and reverse-phase methods for the separation. Protein digests were first loaded on an SCX column. The flow-through peptides from SCX were collected and further loaded on an SAX column. Both columns were eluted by offline pH steps, and the collected fractions were identified by reverse-phase liquid chromatography tandem mass spectrometry. Comprehensive peptide identification was achieved by the Yin-yang MDLC-MS/MS for a 1 mg mouse liver. In total, 14 105 unique peptides were identified with high confidence, including 13 256 unmodified peptides and 849 phosphopeptides with 809 phosphorylated sites. The SCX and SAX in the Yin-Yang system displayed complementary features of binding and separation for peptides. When coupled with reverse-phase liquid chromatography mass spectrometry, the SAX-based method can detect more extremely acidic (pI < 4.0) and phosphorylated peptides, while the SCX-based method detects more relatively basic peptides (pI > 4.0). In total, 134 groups of phosphorylated peptide isoforms were obtained, with common peptide sequences but different phosphorylated states. This unbiased profiling of protein expression and phosphorylation provides a powerful approach to probe protein dynamics, without using any prefractionation and chemical derivation.
学科主题	Biochemistry & Molecular Biology
类目[WOS]	Biochemical Research Methods
关键词[WOS]	LARGE-SCALE ANALYSIS ; PEPTIDE MIXTURES ; PHOSPHOPROTEOMIC ANALYSIS ; SACCHAROMYCES-CEREVISIAE ; AFFINITY-CHROMATOGRAPHY ; PROTEOMIC ANALYSIS ; YEAST PROTEOME ; CELL-CULTURE ; AMINO-ACIDS ; PHOSPHOPEPTIDES
收录类别	SCI
语种	英语
WOS记录号	WOS:000243262600025
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/1508]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Dai, J,Jin, WH,Sheng, QH,et al. Protein phosphorylation and expression profiling by Yin-Yang multidimensional liquid chromatography (Yin-Yang MDLC) mass spectrometry[J]. JOURNAL OF PROTEOME RESEARCH,2007,6(1):250-262.
APA	Dai, J,Jin, WH,Sheng, QH,Shieh, CH,Wu, JR,&Zeng, R.(2007).Protein phosphorylation and expression profiling by Yin-Yang multidimensional liquid chromatography (Yin-Yang MDLC) mass spectrometry.JOURNAL OF PROTEOME RESEARCH,6(1),250-262.
MLA	Dai, J,et al."Protein phosphorylation and expression profiling by Yin-Yang multidimensional liquid chromatography (Yin-Yang MDLC) mass spectrometry".JOURNAL OF PROTEOME RESEARCH 6.1(2007):250-262.