title: Dopamine regulates phosphorylation of VEGF receptor 2 by engaging Src-homology-2-domain-containing protein tyrosine phosphatase 2.
abstract: Vascular endothelial growth factor (VEGF)-induced receptor phosphorylation is the crucial step for initiating downstream signaling pathways that lead to angiogenesis or related pathophysiological outcomes. Our previous studies have shown that the neurotransmitter dopamine could inhibit VEGF-induced phosphorylation of VEGF receptor 2 (VEGFR-2), endothelial cell proliferation, migration, microvascular permeability, and thus, angiogenesis. In this study, we address the mechanism by which VEGFR-2 phosphorylation is regulated by dopamine. Here, we demonstrate that D2 dopamine receptor (D2DR) colocalizes with VEGFR-2 at the cell surface. Dopamine pretreatment increases the translocation and colocalization of Src-homology-2-domain-containing protein tyrosine phosphatase (SHP-2) with D2DR at the cell surface. Dopamine administration leads to increased VEGF-induced phosphorylation of SHP-2 and this increased phosphorylation parallels the increased phosphatase activity of SHP-2. Active SHP-2 then dephosphorylates VEGFR-2 at Y951, Y996 and Y1059, but not Y1175. We also observe that SHP-2 knockdown impairs the dopamine-regulated inhibition of VEGF-induced phosphorylation of VEGFR-2 and, subsequently, Src phosphorylation and migration. Our data establish a novel role for SHP-2 phosphatase in the dopamine-mediated regulation of VEGFR-2 phosphorylation.
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Medicine, Mayo Clinic, Rochester, MN 55905, USA.
authors.forename: Sutapa
authors.initials: S
authors.lastname: Sinha
authors.forename: Pawan Kumar
authors.initials: PK
authors.lastname: Vohra
authors.forename: Resham
authors.initials: R
authors.lastname: Bhattacharya
authors.forename: Shamit
authors.initials: S
authors.lastname: Dutta
authors.forename: Shirshendu
authors.initials: S
authors.lastname: Sinha
authors.forename: Debabrata
authors.initials: D
authors.lastname: Mukhopadhyay
mesh_terms: D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects; D004298:Dopamine / Q000494:pharmacology*; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D055785:Gene Knockdown Techniques; D006801:Humans; D047468:Immunoprecipitation; D008954:Models, Biological; D010766:Phosphorylation / Q000187:drug effects; D019000:Phosphotyrosine / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11 / Q000378:metabolism*; D051076:Proto-Oncogene Proteins c-fyn / Q000378:metabolism; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000378:metabolism; D011720:Pyrazoles / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D019257:Quinpirole / Q000494:pharmacology; D017448:Receptors, Dopamine D2 / Q000378:metabolism; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D014471:Umbilical Veins / Q000166:cytology; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism*
chemical_list: C509621:4-amino-7-phenylpyrazol(3,4-d)pyrimidine; C412373:AG 1879; D011720:Pyrazoles; D011743:Pyrimidines; D017448:Receptors, Dopamine D2; D042461:Vascular Endothelial Growth Factor A; D019257:Quinpirole; D019000:Phosphotyrosine; D040301:Vascular Endothelial Growth Factor Receptor-2; D051076:Proto-Oncogene Proteins c-fyn; D016392:Proto-Oncogene Proteins pp60(c-src); D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11; D004298:Dopamine
references.citation: J Biol Chem. 1999 Apr 30;274(18):12401-7
references.pmid: 10212213
references.citation: J Clin Invest. 2008 Apr;118(4):1380-9
references.pmid: 18340382
references.citation: Trends Biochem Sci. 2003 Jun;28(6):284-93
references.pmid: 12826400
references.citation: J Recept Signal Transduct Res. 2004 Aug;24(3):165-205
references.pmid: 15521361
references.citation: N Engl J Med. 1971 Nov 18;285(21):1182-6
references.pmid: 4938153
references.citation: Nat Med. 2001 May;7(5):569-74
references.pmid: 11329058
references.citation: FASEB J. 1992 Apr;6(7):2413-21
references.pmid: 1348713
references.citation: J Mol Signal. 2008 Mar 31;3:8
references.pmid: 18377662
references.citation: Cancer Treat Rep. 1979 Jun;63(6):991-7
references.pmid: 466656
references.citation: Proc Natl Acad Sci U S A. 2005 Jan 25;102(4):1076-81
references.pmid: 15644447
references.citation: Cell Signal. 2006 Sep;18(9):1447-54
references.pmid: 16442778
references.citation: Science. 1993 Mar 12;259(5101):1611-4
references.pmid: 7681217
references.citation: Prog Clin Biol Res. 1990;354A:317-30
references.pmid: 2247496
references.citation: Adv Neurol. 1990;53:17-29
references.pmid: 1978514
references.citation: Nat Rev Mol Cell Biol. 2006 May;7(5):359-71
references.pmid: 16633338
references.citation: J Mol Signal. 2008 Jul 28;3:14
references.pmid: 18662404
references.citation: J Immunol. 1979 Jan;122(1):166-74
references.pmid: 762415
references.citation: Curr Top Microbiol Immunol. 1999;237:1-30
references.pmid: 9893343
references.citation: J Cell Biol. 2003 May 26;161(4):793-804
references.pmid: 12771128
references.citation: J Biol Chem. 1996 Jun 21;271(25):15160-5
references.pmid: 8663016
references.citation: Mol Cell Biochem. 2004 Sep;264(1-2):51-61
references.pmid: 15544035
references.citation: Blood. 2005 Mar 1;105(5):1970-6
references.pmid: 15507525
references.citation: Biochem Soc Trans. 2003 Dec;31(Pt 6):1171-7
references.pmid: 14641020
references.citation: Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):7335-9
references.pmid: 8041791
references.citation: J Biol Chem. 2001 Aug 31;276(35):32714-9
references.pmid: 11435426
references.citation: Curr Top Microbiol Immunol. 1999;237:97-132
references.pmid: 9893348
references.citation: Am J Physiol Heart Circ Physiol. 2004 Oct;287(4):H1554-60
references.pmid: 15371263
references.citation: Nat Med. 1995 Jan;1(1):27-31
references.pmid: 7584949
references.citation: Curr Opin Neurobiol. 1997 Oct;7(5):701-7
references.pmid: 9384552
references.citation: Endocrine. 2000 Jun;12(3):237-41
references.pmid: 10963043
references.citation: Ann N Y Acad Sci. 1997 Apr 15;811:311-22; 322-4
references.pmid: 9186607
references.citation: Nat Med. 2003 Jun;9(6):669-76
references.pmid: 12778165
references.citation: J Invest Dermatol. 1978 Aug;71(2):163-4
references.pmid: 681787
references.citation: BMB Rep. 2008 Apr 30;41(4):278-86
references.pmid: 18452647
references.citation: Nature. 1997 Apr 17;386(6626):671-4
references.pmid: 9109485
references.citation: Circ Res. 2006 Jan 6;98(1):45-54
references.pmid: 16339483
references.citation: Crit Care Med. 2008 Jan;36(1):296-327
references.pmid: 18158437
references.citation: J Biochem Mol Biol. 2006 Sep 30;39(5):469-78
references.pmid: 17002866
references.citation: Cell. 2003 Jul 25;114(2):171-80
references.pmid: 12887919
references.citation: Mol Cell. 2001 Oct;8(4):759-69
references.pmid: 11684012
references.citation: EMBO J. 2005 Jul 6;24(13):2342-53
references.pmid: 15962004
references.citation: EMBO J. 2001 Jun 1;20(11):2768-78
references.pmid: 11387210
references.citation: Nat Med. 2003 Jun;9(6):653-60
references.pmid: 12778163
references.citation: Clin Cancer Res. 2008 Apr 15;14(8):2502-10
references.pmid: 18413843
references.citation: Annu Rev Neurosci. 1999;22:123-44
references.pmid: 10202534