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Small GTP-binding proteins in human endothelial cells
Authors:Hubert P. J. C. De Leeuw,Pauline M. Koster,Jero Calafat,Hans Janssen,Anton-Jan Van Zonneveld,Jan A. Van Mourik,&   Jan Voorberg
Affiliation:Department of Blood Coagulation, CLB, Sanquin Blood Supply Foundation, Amsterdam; Department of Cell Biology, Netherlands Cancer Institute, Amsterdam; Department of Biochemistry, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands
Abstract:Small GTP-binding proteins of the Ras superfamily control an extensive number of intracellular events by alternating between GDP- and GTP-bound conformation. The presence of members of this protein family was examined in human umbilical vein endothelial cells employing RT-PCR. Sequence analysis of 215 cDNA clones revealed the presence of a total of 28 different partial cDNAs encoding small GTP-binding proteins. Two sequences corresponded to novel isoforms of Rab2 and Rab9. In addition, human analogues of Rab4b, Rab7, Rab9, Rab14 and Rab15 were identified. Besides Rab proteins, members of other subfamilies were detected as well. As a first step towards elucidation of the function of the different small GTP-binding proteins identified we have isolated full length cDNA corresponding to Rab30 from a human endothelial cell cDNA library. In order to assess the subcellular localization of Rab30, we expressed epitope-tagged Rab30 cDNA in monkey kidney COS-1 cells. Immunoelectron-microscopy of transfected COS-1 cells indicated that Rab30 is associated with Golgi stacks.
Keywords:endothelial cells    GTP-binding proteins    Rab30    Golgi apparatus
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