title: Pigment epithelium-derived factor binds to cell-surface F(1)-ATP synthase.
abstract: Pigment epithelium-derived factor (PEDF), a potent blocker of angiogenesis in vivo, and of endothelial cell migration and tubule formation, binds with high affinity to an as yet unknown protein on the surfaces of endothelial cells. Given that protein fingerprinting suggested a match of a approximately 60 kDa PEDF-binding protein in bovine retina with Bos taurus F(1)-ATP synthase beta-subunit, and that F(1)F(o)-ATP synthase components have been identified recently as cell-surface receptors, we examined the direct binding of PEDF to F(1). Size-exclusion ultrafiltration assays showed that recombinant human PEDF formed a complex with recombinant yeast F(1). Real-time binding as determined by surface plasmon resonance demonstrated that yeast F(1) interacted specifically and reversibly with human PEDF. Kinetic evaluations revealed high binding affinity for PEDF, in agreement with PEDF affinities for endothelial cell surfaces. PEDF blocked interactions between F(1) and angiostatin, another antiangiogenic factor, suggesting overlapping PEDF-binding and angiostatin-binding sites on F(1). Surfaces of endothelial cells exhibited affinity for PEDF-binding proteins of approximately 60 kDa. Antibodies to F(1)beta-subunit specifically captured PEDF-binding components in endothelial plasma membranes. The extracellular ATP synthesis activity of endothelial cells was examined in the presence of PEDF. PEDF significantly reduced the amount of extracellular ATP produced by endothelial cells, in agreement with direct interactions between cell-surface ATP synthase and PEDF. In addition to demonstrating that PEDF binds to cell-surface F(1), these results show that PEDF is a ligand for endothelial cell-surface F(1)F(o)-ATP synthase. They suggest that PEDF-mediated inhibition of ATP synthase may form part of the biochemical mechanisms by which PEDF exerts its antiangiogenic activity.
authors.affiliation: Section of Protein Structure and Function, Laboratory of Retinal Cell and Molecular Biology, National Eye Institute, NIH, Bethesda, MD, USA.
authors.forename: Luigi
authors.initials: L
authors.lastname: Notari
authors.forename: Naokatu
authors.initials: N
authors.lastname: Arakaki
authors.forename: David
authors.initials: D
authors.lastname: Mueller
authors.forename: Scott
authors.initials: S
authors.lastname: Meier
authors.forename: Juan
authors.initials: J
authors.lastname: Amaral
authors.forename: S P
authors.initials: SP
authors.lastname: Becerra
mesh_terms: D000255:Adenosine Triphosphate / Q000096:biosynthesis; D000818:Animals; D001667:Binding, Competitive; D002417:Cattle; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000378:metabolism; D005136:Eye Proteins / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D009414:Nerve Growth Factors / Q000378:metabolism*; D011485:Protein Binding; D021122:Protein Subunits / Q000378:metabolism; D006180:Proton-Translocating ATPases / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D015843:Serpins / Q000378:metabolism*; D020349:Surface Plasmon Resonance; D000099174:Pigment Epithelium-Derived Factor
chemical_list: D000255:Adenosine Triphosphate; D005136:Eye Proteins; D009414:Nerve Growth Factors; D021122:Protein Subunits; D006180:Proton-Translocating ATPases; D015843:Serpins; D000099174:Pigment Epithelium-Derived Factor
references.citation: J Biol Chem. 2005 Jan 21;280(3):1740-5
references.pmid: 15528193
references.citation: Protein Expr Purif. 2004 Oct;37(2):479-85
references.pmid: 15358374
references.citation: Invest Ophthalmol Vis Sci. 2005 Aug;46(8):2736-47
references.pmid: 16043845
references.citation: Cancer Res. 2005 Jun 15;65(12):5144-52
references.pmid: 15958558
references.citation: Proc Natl Acad Sci U S A. 2001 Feb 27;98(5):2593-7
references.pmid: 11226284
references.citation: J Biol Chem. 2006 Dec 8;281(49):38022-37
references.pmid: 17032652
references.citation: J Biol Chem. 2002 Nov 22;277(47):45400-7
references.pmid: 12237317
references.citation: Proc Natl Acad Sci U S A. 1999 Mar 16;96(6):2811-6
references.pmid: 10077593
references.citation: J Cell Biochem. 2005 Oct 1;96(2):242-61
references.pmid: 16094651
references.citation: Trends Mol Med. 2006 Oct;12(10):497-502
references.pmid: 16962374
references.citation: Diabetes. 2001 Dec;50(12):2641-5
references.pmid: 11723044
references.citation: Trends Mol Med. 2002 Jul;8(7):330-4
references.pmid: 12114112
references.citation: Invest Ophthalmol Vis Sci. 2001 Dec;42(13):3287-93
references.pmid: 11726635
references.citation: Invest Ophthalmol Vis Sci. 2010 Mar;51(3):1318-26
references.pmid: 19850839
references.citation: Protein Sci. 1996 Dec;5(12):2575-82
references.pmid: 8976566
references.citation: J Cell Physiol. 2001 Aug;188(2):253-63
references.pmid: 11424092
references.citation: J Cell Sci. 2001 Dec;114(Pt 24):4421-8
references.pmid: 11792807
references.citation: Proc Natl Acad Sci U S A. 2007 Aug 21;104(34):13632-7
references.pmid: 17698806
references.citation: J Neurosci. 2002 Nov 1;22(21):9378-86
references.pmid: 12417663
references.citation: Curr Opin Lipidol. 2006 Jun;17(3):279-84
references.pmid: 16680033
references.citation: Biochem Biophys Res Commun. 2004 Sep 3;321(4):788-94
references.pmid: 15358096
references.citation: Anticancer Res. 2000 Sep-Oct;20(5A):2923-30
references.pmid: 11062702
references.citation: J Mol Cell Cardiol. 2004 Aug;37(2):497-506
references.pmid: 15276019
references.citation: J Biol Chem. 1999 Oct 29;274(44):31605-12
references.pmid: 10531367
references.citation: Nat Med. 2003 Jun;9(6):774-80
references.pmid: 12740569
references.citation: Neurobiol Dis. 1999 Dec;6(6):523-32
references.pmid: 10600408
references.citation: Exp Eye Res. 2006 May;82(5):739-40
references.pmid: 16364294
references.citation: Proc Natl Acad Sci U S A. 2001 Jun 5;98(12):6656-61
references.pmid: 11381144
references.citation: Prog Retin Eye Res. 2004 Sep;23(5):561-77
references.pmid: 15302351
references.citation: Cancer Res. 2004 May 15;64(10):3679-86
references.pmid: 15150128
references.citation: Biochemistry. 1998 Jul 28;37(30):10643-52
references.pmid: 9692954
references.citation: Invest Ophthalmol Vis Sci. 1996 Sep;37(10):1984-93
references.pmid: 8814138
references.citation: Protein Expr Purif. 1995 Aug;6(4):447-56
references.pmid: 8527930
references.citation: Invest Ophthalmol Vis Sci. 2002 May;43(5):1574-80
references.pmid: 11980876
references.citation: Mol Ther. 2003 Jul;8(1):72-9
references.pmid: 12842430
references.citation: Invest Ophthalmol Vis Sci. 2002 Jun;43(6):1994-2000
references.pmid: 12037010
references.citation: Mol Cancer Res. 2003 Nov;1(13):931-9
references.pmid: 14638865
references.citation: Invest Ophthalmol Vis Sci. 2003 Oct;44(10):4497-504
references.pmid: 14507898
references.citation: Science. 1999 Jul 9;285(5425):245-8
references.pmid: 10398599
references.citation: Proc Natl Acad Sci U S A. 2002 Jun 25;99(13):8909-14
references.pmid: 12072560
references.citation: Cancer Res. 2006 Jan 15;66(2):875-82
references.pmid: 16424020