title: Phosphotyrosine enrichment identifies focal adhesion kinase and other tyrosine kinases for targeting in canine hemangiosarcoma.
abstract: Canine hemangiosarcoma (HSA) is an endothelial cell malignancy driven, in part, by activating mutations in receptor and non-receptor tyrosine kinases. Proteomics, Western blots and a tyrosine kinase inhibitor were used to elucidate activating mechanisms in HSA cell lines. Phosphotyrosine peptides from focal adhesion kinase (FAK) STAT3, Lyn, Fyn and other signal transduction kinases were identified by mass spectrometry. FAK was constitutively activated at tyrosine 397, the autophosphorylation site, and this was reversible with high concentrations of a FAK inhibitor. FAK inhibitor-14 suppressed migration and phosphorylation of FAK tyrosine 397 and tyrosines 576/577 and was cytotoxic to HSA cells suggesting FAK signalling may be an important contributor to canine HSA survival.
authors.affiliation: Department of Clinical Sciences, Oregon State University, Corvallis, OR 97331, USA.
authors.forename: K
authors.initials: K
authors.lastname: Marley
authors.forename: C S
authors.initials: CS
authors.lastname: Maier
authors.forename: S C
authors.initials: SC
authors.lastname: Helfand
mesh_terms: D000818:Animals; D015153:Blotting, Western / Q000662:veterinary; D045744:Cell Line, Tumor; D004283:Dog Diseases / Q000188:drug therapy / Q000201:enzymology*; D004285:Dogs; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006394:Hemangiosarcoma / Q000188:drug therapy / Q000201:enzymology / Q000662:veterinary*; D013058:Mass Spectrometry / Q000662:veterinary; D009383:Neoplasms, Vascular Tissue / Q000188:drug therapy / Q000201:enzymology / Q000662:veterinary*; D019000:Phosphotyrosine / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D051076:Proto-Oncogene Proteins c-fyn / Q000187:drug effects / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000187:drug effects / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D019061:src-Family Kinases / Q000187:drug effects / Q000378:metabolism
chemical_list: D050796:STAT3 Transcription Factor; D019000:Phosphotyrosine; D011505:Protein-Tyrosine Kinases; D051416:Focal Adhesion Protein-Tyrosine Kinases; D051076:Proto-Oncogene Proteins c-fyn; C060902:lyn protein-tyrosine kinase; D019061:src-Family Kinases