title: The enzymatic activity of the VEGFR2 receptor for the biosynthesis of dinucleoside polyphosphates.
abstract: The group of dinucleoside polyphosphates encompasses a large number of molecules consisting of two nucleosides which are connected by a phosphate chain of variable length. While the receptors activated by dinucleoside polyphosphates as well as their degradation have been studied in detail, its biosynthesis has not been elucidated so far. Since endothelial cells released the dinucleoside polyphosphate uridine adenosine tetraphosphate (Up4A), we tested cytosolic proteins of human endothelial cells obtained from dermal vessels elicited for enzymatic activity. When incubated with ADP and UDP, these cells showed increasing concentrations of Up4A. The underlying enzyme was isolated by chromatography and the mass spectrometric analysis revealed that the enzymatic activity was caused by the vascular endothelial growth factor receptor 2 (VEGFR2). Since VEGFR2 but neither VEGFR1 nor VEGFR3 were capable to synthesise dinucleoside polyphosphates, Tyr-1175 of VEGFR2 is most likely essential for the enzymatic activity of interest. Further, VEGFR2-containing cells like HepG2, THP-1 and RAW264.7 were capable of synthesising dinucleoside polyphosphates. VEGFR2-transfected HEK 293T/17 but not native HEK 293T/17 cells synthesised dinucleoside polyphosphates in vivo too. The simultaneous biosynthesis of dinucleoside polyphosphates could amplify the response to VEGF, since dinucleoside polyphosphates induce cellular growth via P2Y purinergic receptors. Thus the biosynthesis of dinucleoside polyphosphates by VEGFR2 may enhance the proliferative response to VEGF. Given that VEGFR2 is primarily expressed in endothelial cells, the biosynthesis of dinucleoside polyphosphates is mainly located in the vascular system. Since the vasculature is also the main site of action of dinucleoside polyphosphates, activating vascular purinoceptors, blood vessels appear as an autocrine system with respect to dinucleoside polyphosphates. We conclude that VEGFR2 receptor is capable of synthesising dinucleoside polyphosphates. These mediators may modulate the effects of VEGFR2 due to their proliferative effects.
authors: Charité-Universitaetsmedizin Berlin, Medizinische Klinik IV, Hindenburgdamm 30, D-12200, Berlin, Germany. Joachim.Jankowski@charite.de
authors: Vera
authors: V
authors: Jankowski
authors: Anna
authors: A
authors: Schulz
authors: Axel
authors: A
authors: Kretschmer
authors: Harald
authors: H
authors: Mischak
authors: Falko
authors: F
authors: Boehringer
authors: Markus
authors: M
authors: van der Giet
authors: Doreen
authors: D
authors: Janke
authors: Mirjam
authors: M
authors: Schuchardt
authors: Ralf
authors: R
authors: Herwig
authors: Walter
authors: W
authors: Zidek
authors: Joachim
authors: J
authors: Jankowski
mesh_terms: D000244:Adenosine Diphosphate / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D015226:Dinucleoside Phosphates / Q000096:biosynthesis*; D057809:HEK293 Cells; D056945:Hep G2 Cells; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D051379:Mice; D034741:RNA, Small Interfering / Q000235:genetics; D014530:Uridine Diphosphate / Q000494:pharmacology; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism*; D040321:Vascular Endothelial Growth Factor Receptor-3 / Q000378:metabolism
chemical_list: D015226:Dinucleoside Phosphates; D034741:RNA, Small Interfering; C529314:uridine adenosine tetraphosphate; D014530:Uridine Diphosphate; D000244:Adenosine Diphosphate; D040281:Vascular Endothelial Growth Factor Receptor-1; D040301:Vascular Endothelial Growth Factor Receptor-2; D040321:Vascular Endothelial Growth Factor Receptor-3
references: J Cardiovasc Pharmacol. 2011 Dec;58(6):654-62
references: 21885991
references: Reproduction. 2009 Dec;138(6):883-93
references: 19755482
references: Clin Cancer Res. 2011 Jul 1;17(13):4439-50
references: 21622720
references: Electrophoresis. 1999 Dec;20(18):3551-67
references: 10612281
references: Biochimie. 2001 Mar-Apr;83(3-4):351-6
references: 11295496
references: Nature. 1994 Jan 13;367(6459):186-8
references: 8114917
references: Pharmacol Res. 2012 Jan;65(1):81-90
references: 21933714
references: Arterioscler Thromb Vasc Biol. 2007 Feb;27(2):297-302
references: 17138938
references: Adv Pharmacol Sci. 2011;2011:435132
references: 22110488
references: Genes Dev. 2000 Oct 15;14(20):2610-22
references: 11040215
references: J Biol Chem. 2004 May 21;279(21):22267-75
references: 15026417
references: Arterioscler Thromb Vasc Biol. 2007 Aug;27(8):1776-81
references: 17569882
references: Anal Chem. 2001 Feb 1;73(3):434-8
references: 11217742
references: J Med Chem. 2012 Jan 12;55(1):437-48
references: 22107038
references: Bioinformatics. 2007 Nov 1;23(21):2947-8
references: 17846036
references: Cancer Res. 2006 Dec 15;66(24):11851-8
references: 17178882
references: Cell Res. 2009 May;19(5):651-64
references: 19399029
references: Am J Physiol Lung Cell Mol Physiol. 2009 Oct;297(4):L758-66
references: 19592459
references: Proc Natl Acad Sci U S A. 1976 Nov;73(11):3984-8
references: 1069282
references: Semin Dial. 2009 Jul-Aug;22(4):396-9
references: 19708989
references: Biochem Biophys Res Commun. 2012 Jan 20;417(3):1035-40
references: 22214933
references: Pharmacol Res. 2013 Jan;67(1):10-7
references: 23063485
references: J Cell Sci. 2009 Sep 15;122(Pt 18):3303-11
references: 19706681
references: Cancer Res. 2008 Jun 1;68(11):4340-6
references: 18519694
references: Biosci Rep. 2008 Aug;28(4):189-94
references: 18576946
references: Nat Med. 2005 Feb;11(2):223-7
references: 15665829
references: Nucleic Acids Res. 2011 Jan;39(Database issue):D214-9
references: 21051339
references: J Biol Chem. 1999 Aug 20;274(34):23926-31
references: 10446159
references: Exp Hematol. 2002 Jul;30(7):767-73
references: 12135675
references: Proc Natl Acad Sci U S A. 2004 Sep 14;101(37):13489-94
references: 15342917
references: Br J Pharmacol. 2009 Aug;157(7):1142-53
references: 19563527
references: Biochem J. 1982 Dec 15;208(3):737-42
references: 6299279
references: Biochem Biophys Res Commun. 1983 Aug 30;115(1):253-60
references: 6311204