title: SH2 Domains as Affinity Reagents for Phosphotyrosine Protein Enrichment and Proteomic Analysis.
abstract: Dynamic tyrosine phosphorylation is a key molecular modulation for many signal transduction events. Because of their low abundance and dynamic nature in cells, the detection and enrichment of phosphotyrosine proteins has long relied on specific antibodies, such as 4G10 and P-Tyr-100. Another well-established approach for phosphotyrosine proteins recognition and enrichment is by their specific binding domains, such as Src homology 2 (SH2) domains. In this chapter, we describe a typical analytical approach for purifying specific SH2 domains, enriching specific phosphotyrosine proteins from activated cells, mass spectrometry analysis, and related data analysis.
authors.affiliation: Department of Chemistry, South University of Science and Technology of China, Shenzhen, 518055, People's Republic of China.
authors.forename: Mi
authors.initials: M
authors.lastname: Ke
authors.affiliation: Department of Chemistry, South University of Science and Technology of China, Shenzhen, 518055, People's Republic of China.
authors.forename: Bizhu
authors.initials: B
authors.lastname: Chu
authors.affiliation: Department of Chemistry, South University of Science and Technology of China, Shenzhen, 518055, People's Republic of China.|Materials Characterization and Preparation Center, South University of Science and Technology of China, Shenzhen, 518055, People's Republic of China.
authors.forename: Lin
authors.initials: L
authors.lastname: Lin
authors.affiliation: Department of Chemistry, South University of Science and Technology of China, Shenzhen, 518055, People's Republic of China. tian.rj@sustc.edu.cn.|Shenzhen Key Laboratory of Cell Microenvironment, South University of Science and Technology of China, Shenzhen, 518055, People's Republic of China. tian.rj@sustc.edu.cn.
authors.forename: Ruijun
authors.initials: R
authors.lastname: Tian
mesh_terms: D002853:Chromatography, Liquid; D006801:Humans; D019169:Jurkat Cells; D013058:Mass Spectrometry; D010750:Phosphoproteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D019000:Phosphotyrosine / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D040901:Proteomics / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D015398:Signal Transduction; D057188:Workflow; D018909:src Homology Domains
chemical_list: D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; D019000:Phosphotyrosine
keywords: Affinity reagent; LC-MS; Phosphotyrosine; Proteomics; SH2 domain; Signal transduction