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Analysis of the Subcellular Phosphoproteome Using a Novel Phosphoproteomic Reactor
Zhou, Houjiang1,2,3,5; Elisma, Fred2,3; Denis, Nicholas J.2,3; Wright, Theodore G.2,3; Tian, Ruijun2,3; Zhou, Hu2,3; Hou, Weimin2,4; Zou, Hanfa1,5; Figeys, Daniel2,3,4; Zou HF(邹汉法); Daniel Figeys
KeywordPhosphoproteomics Proteomic Reactor Subcellular Technology
Source PublicationJOURNAL OF PROTEOME RESEARCH
2010-03-01
ISSN1535-3893
DOI10.1021/pr900767j
Volume9Issue:3Pages:1279-1288
Indexed BySCI
SubtypeArticle
Department18
Funding Project1809
Contribution Rank3;1
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
WOS SubjectBiochemical Research Methods
WOS Research AreaBiochemistry & Molecular Biology
WOS KeywordION AFFINITY-CHROMATOGRAPHY ; HIGHLY SELECTIVE ENRICHMENT ; CYTOSCAPE PLUG-IN ; MASS-SPECTROMETRY ; PHOSPHORYLATED PEPTIDES ; PROTEOMIC REACTOR ; QUANTITATIVE PHOSPHOPROTEOMICS ; PHOSPHOPEPTIDE ENRICHMENT ; ORGANELLE PROTEOMICS ; TITANIUM-DIOXIDE
AbstractProtein phosphorylation is an important post-translational modification involved in the regulation of many cellular processes. Mass spectrometry has been successfully used to identify protein phosphorylation in specific pathways and for global phosphoproteomic analysis. However, phosphoproteomics approaches do not evaluate the subcellular localization of the phosphorylated forms of proteins, which is an important factor for understanding the roles of protein phosphorylation on a global scale. The in-depth mapping of protein phosphorylation at the subcellular level necessitates the development of new methods capable of specifically and efficiently enriching phosphopeptides from highly complex samples. Here, we report a novel microfluidic device called the phosphoproteomic reactor that combines efficient processing of proteins followed by phosphopeptide enrichment by Ti-IMAC. To illustrate the potential of this novel technology, we mapped the phosphoproteins in subcellular organelles of liver cells. Fifteen subcellular fractions from liver cell cultures were processed on the phosphoproteomic reactor in combination with nano-LC-MS/MS analysis. We identified thousands of phosphorylation sites in over 600 phosphoproteins in different organelles using minute amounts of starting material. Overall, this approach provides a new avenue for studying the phosphoproteome of the subcellular organelles.
Language英语
URL查看原文
WOS IDWOS:000275088100011
Citation statistics
Cited Times:22[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/103065
Collection中国科学院大连化学物理研究所
Corresponding AuthorZou HF(邹汉法); Daniel Figeys
Affiliation1.Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Univ Ottawa, Ottawa Inst Syst Biol, Ottawa, ON K1H 8M5, Canada
3.Univ Ottawa, Dept Biochem Microbiol & Immunol, Fac Med, Ottawa, ON K1H 8M5, Canada
4.Univ Ottawa, Dept Chem, Ottawa, ON K1H 8M5, Canada
5.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Dalian 116023, Peoples R China
Recommended Citation
GB/T 7714
Zhou, Houjiang,Elisma, Fred,Denis, Nicholas J.,et al. Analysis of the Subcellular Phosphoproteome Using a Novel Phosphoproteomic Reactor[J]. JOURNAL OF PROTEOME RESEARCH,2010,9(3):1279-1288.
APA Zhou, Houjiang.,Elisma, Fred.,Denis, Nicholas J..,Wright, Theodore G..,Tian, Ruijun.,...&Daniel Figeys.(2010).Analysis of the Subcellular Phosphoproteome Using a Novel Phosphoproteomic Reactor.JOURNAL OF PROTEOME RESEARCH,9(3),1279-1288.
MLA Zhou, Houjiang,et al."Analysis of the Subcellular Phosphoproteome Using a Novel Phosphoproteomic Reactor".JOURNAL OF PROTEOME RESEARCH 9.3(2010):1279-1288.
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