Comprehensive Profiling of Phosphopeptides Based on Anion Exchange Followed by Flow-Through Enrichment with Titanium Dioxide (AFET)

	Comprehensive Profiling of Phosphopeptides Based on Anion Exchange Followed by Flow-Through Enrichment with Titanium Dioxide (AFET)
	Nie, S; Dai, J; Ning, ZB; Cao, XJ; Sheng, QH; Zeng, R
刊名	JOURNAL OF PROTEOME RESEARCH
	2010
卷号	9 期号:9 页码:4585-4594
关键词	Phosphopeptide anion exchange chromatography titanium dioxide phosphoproteomics
通讯作者	Zeng, R (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, Key Lab Syst Biol, 320 Yue Yang Rd, Shanghai 200031, Peoples R China.,zr@sibs.ac.cn
英文摘要	For large-scale analysis of phosphorylation at proteome-wide scale, a variety of affinity-based strategies have been developed to enrich phosphopeptide. Because each method differed in their specificity of isolation, the global and unbiased enrichment of phosphopeptides remains a major technical challenge in phosphoproteomics. In the present work, we demostrate that the phosphopeptide enrichment method based on an online continuous pH gradient in a strong anion exchange column (SAX method) is highly complementary to the method based on titanium dioxide (TiO(2)) affinity enrichment. Moreover, we found that the flow-through fraction of either SAX or SCX is very phosphopeptide-rich, which necessitates further analysis by complementary method. Here, we developed a comprehensive phosphopeptides profiling strategy based on anion exchange followed by flow-through enrichment by TiO(2) (AFET). In this strategy, SAX method was used as the first separation/enrichment step, which was online coupled with LC-MS/MS. The phosphopeptides in the SAX flow-through fraction were further enriched with TiO(2). As a result, a more comprehensive, less biased phosphoproteome was aquired. Careful comparison of four different combination strategies reveal that the AFET method showed the advantages of more identified phosphopeptides, less mass spectrometry analysis time, as well as simple and automatic process step. It is well-suited for robust and reproducible phosphoproteomics, especially in the case of small amounts of sample.
学科主题	Biochemistry & Molecular Biology
类目[WOS]	Biochemical Research Methods
关键词[WOS]	CALCIUM-PHOSPHATE PRECIPITATION ; METAL-OXIDE CHROMATOGRAPHY ; MASS-SPECTROMETRY ; PHOSPHOPROTEOME ANALYSIS ; QUANTITATIVE PHOSPHOPROTEOMICS ; PHOSPHORYLATED PEPTIDES ; PROTEIN-PHOSPHORYLATION ; AFFINITY-CHROMATOGRAPHY ; DROSOPHILA-MELANOGASTER ; SIGNALING PATHWAY
收录类别	SCI
语种	英语
WOS记录号	WOS:000281443700024
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/1050]
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	Nie, S,Dai, J,Ning, ZB,et al. Comprehensive Profiling of Phosphopeptides Based on Anion Exchange Followed by Flow-Through Enrichment with Titanium Dioxide (AFET)[J]. JOURNAL OF PROTEOME RESEARCH,2010,9(9):4585-4594.
APA	Nie, S,Dai, J,Ning, ZB,Cao, XJ,Sheng, QH,&Zeng, R.(2010).Comprehensive Profiling of Phosphopeptides Based on Anion Exchange Followed by Flow-Through Enrichment with Titanium Dioxide (AFET).JOURNAL OF PROTEOME RESEARCH,9(9),4585-4594.
MLA	Nie, S,et al."Comprehensive Profiling of Phosphopeptides Based on Anion Exchange Followed by Flow-Through Enrichment with Titanium Dioxide (AFET)".JOURNAL OF PROTEOME RESEARCH 9.9(2010):4585-4594.