title: Toward a comprehensive characterization of the phosphotyrosine proteome.
abstract: Tyrosine phosphorylation (pTyr) regulates important cell functions and plays a key role in carcinogenesis. The purpose of this study was to perform a comprehensive study of the phosphotyrosine proteome. Immunoaffinity enriched pTyr proteins and peptides from K562 leukemia cells were analyzed with high-resolving liquid chromatography mass spectrometry. Two different antibodies selective for the pTyr modification were used in repeated enrichments to identify as many pTyr peptides as possible. Stringent verification of putative pTyr sites was performed to assure high reliability in the subsequent biological interpretation of the data. Identified pTyr proteins were subjected to pathway analysis by using different analytical tools. In total, 294 pTyr peptides belonging to 217 pTyr proteins were identified, 15 of which had not previously been reported to be modified by pTyr. The pTyr proteins were clustered in six major groups based on the biological functions "cellular signaling", "cell motility and shape", "cell cycle process", "transport", "RNA processing" and "protein processing". The pTyr proteins were mainly positioned in the following cellular compartments: cytoplasm, cytoskeleton, nucleus and ribonucleoprotein complexes. An interesting finding was that many proteins were related to RNA processing and were found to be heterogeneous nuclear ribonucleoproteins. Also, more than half of the novel pTyr proteins were localized to the nucleus, of which three (PBX2, TEAD1 and DIDO1) were classified as transcription factors and two (CENPC1 and MAD2L1) are associated with cell division control.
authors: Department of Immunology, Genetics and Pathology, Rudbeck Laboratory, Uppsala University, SE-751 85 Uppsala, Sweden. sara.lind@igp.uu.se
authors: Sara
authors: S
authors: Bergström Lind
authors: Konstantin A
authors: KA
authors: Artemenko
authors: Lioudmila
authors: L
authors: Elfineh
authors: Corina
authors: C
authors: Mayrhofer
authors: Roman A
authors: RA
authors: Zubarev
authors: Jonas
authors: J
authors: Bergquist
authors: Ulf
authors: U
authors: Pettersson
mesh_terms: D000906:Antibodies / Q000276:immunology; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D020014:K562 Cells; D013058:Mass Spectrometry / Q000379:methods; D008954:Models, Biological; D010455:Peptides / Q000032:analysis / Q000302:isolation & purification; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000032:analysis*; D020543:Proteome / Q000032:analysis*
chemical_list: D000906:Antibodies; D010455:Peptides; D020543:Proteome; D019000:Phosphotyrosine