Sck is expressed in endothelial cells and participates in vascular endothelial growth factor-induced signaling

Sck, a member of the Shc family of cell signaling proteins, has only been studied in neuronal cells, though previous studies have demonstrated its expression in tissues other than brain. Using RT-PCR and RNase protection assays, we detected Sck mRNA expression in endothelial cells, and Sck protein w...

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Bibliographic Details
Published in:	Oncogene Vol. 21; no. 41; pp. 6307 - 6316
Main Authors:	RATCLIFFE, Kirsty E, QI TAO, YAVUZ, Burju, STOLETOV, Konstantin V, SPRING, Simone C, TERMAN, Bruce I
Format:	Journal Article
Language:	English
Published:	Basingstoke Nature Publishing 12-09-2002 Nature Publishing Group
Subjects:	Adaptor Proteins, Signal Transducing Angiogenesis Animals Aorta Biological and medical sciences Cardiology Cell physiology Cell signaling Endothelial cells Endothelial Growth Factors - metabolism Endothelium, Vascular - physiology Fundamental and applied biological sciences. Psychology Gene expression Humans Immunohistochemistry Immunoprecipitation Internalization Kinases Lymphokines - metabolism MAP kinase Medicine Mice Molecular and cellular biology Monoclonal antibodies Organ Specificity Peptides Phosphorylation Phosphotyrosine Platelet Endothelial Cell Adhesion Molecule-1 - metabolism Proteins Proteins - metabolism Receptor Protein-Tyrosine Kinases - metabolism Receptors, Growth Factor - metabolism Receptors, Vascular Endothelial Growth Factor Retina Ribonuclease RNA, Messenger - analysis Shc Signaling Adaptor Proteins Signal Transduction Src Homology 2 Domain-Containing, Transforming Protein 2 Swine Tyrosine Umbilical vein Vascular endothelial growth factor Vascular Endothelial Growth Factor A Vascular Endothelial Growth Factors Western blotting New York United States > US Endothelial cell Biochemical analysis Enzyme Family study Esterases Protein Tissue Vascular endothelium growth factor Messenger RNA Immunoblotting assay Hydrolases Pancreatic ribonuclease Protection Reverse transcription polymerase chain reaction Polyclonal immunoglobulin Brain (vertebrata)
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Summary:	Sck, a member of the Shc family of cell signaling proteins, has only been studied in neuronal cells, though previous studies have demonstrated its expression in tissues other than brain. Using RT-PCR and RNase protection assays, we detected Sck mRNA expression in endothelial cells, and Sck protein was detected by Western blotting using polyclonal and monoclonal antibodies targeting the Sck CH1 domain. Immunohistochemistry protocols demonstrate that Sck is expressed in KDR and PECAM positive cells found in the mouse retina, mouse heart and human umbilical chord. Treatment of human umbilical vein endothelial (HUVE) cells with vascular endothelial growth factor (VEGF) leads to the recruitment of Sck to the KDR VEGF receptor and an enhanced Sck tyrosine phosphorylation. Sck is recruited to KDR tyrosine 1175, as co-immunoprecipitation of KDR and Sck is not observed in VEGF-treated porcine aortic endothelial cells expressing a receptor mutated at this autophosphorylation site. The Sck and Shc SH2 domains, and not the PTB domain, mediates its interactions with KDR, as recombinant Sck SH2 domain binds to a tyrosine phosphorylated KDR 1175-derived synthetic peptide, but not to a peptide synthesized without tyrosine phosphate. Recombinant PLCgamma SH2 domain also interacts with the phosphotyrosine 1175 containing peptide. VEGF-induced MAPK activation is dependent upon PLCgamma activity, and chimeric proteins consisting of the Shc or Sck SH2 domains fused with a cellular internalization sequence attenuated this activation. Taken together, these results demonstrate that Sck is expressed in vascular endothelial cells, and participates in VEGF-induced signal transduction.
Bibliography:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2
ISSN:	0950-9232 1476-5594
DOI:	10.1038/sj.onc.1205781