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The enzymatic activity of the VEGFR2 receptor for the biosynthesis of dinucleoside polyphosphates

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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.

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Acknowledgements

This study was supported by a grant of the German Research Foundation (DFG, Ja-972 /11-1/2), a grant from the Federal Ministry of Education and Research (01GR080701/09), grant FP7-HEALTH-2009-2.4.5-2 to “SysKid” (grant agreement 241544), “Mascara” (grant agreement 278249) from the European Union and by the Peter und Traudl Engelhorn Stiftung. We thank B. Egbers and D. Jacobi for excellent technical assistance.

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Authors and Affiliations

  1. Charité-Universitaetsmedizin Berlin, Medizinische Klinik IV, Hindenburgdamm 30, D-12200, Berlin, Germany

    Vera Jankowski, Anna Schulz, Falko Boehringer, Markus van der Giet, Doreen Janke, Mirjam Schuchardt, Walter Zidek & Joachim Jankowski

  2. Bayer Pharma AG, Wuppertal, Germany

    Axel Kretschmer

  3. Biomarkers and Systems Medicine, University of Glasgow, Proteomic, Glasgow, UK

    Harald Mischak

  4. Department Vertebrate Genomics, Max Planck Institute for Molecular Genetics, 14195, Berlin, Germany

    Ralf Herwig

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  1. Vera Jankowski
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Jankowski, V., Schulz, A., Kretschmer, A. et al. The enzymatic activity of the VEGFR2 receptor for the biosynthesis of dinucleoside polyphosphates. J Mol Med 91, 1095–1107 (2013). https://doi.org/10.1007/s00109-013-1036-y

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  • DOI: https://doi.org/10.1007/s00109-013-1036-y

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