title: Kindlin-2 regulates hemostasis by controlling endothelial cell-surface expression of ADP/AMP catabolic enzymes via a clathrin-dependent mechanism.
abstract: Kindlin-2, a widely distributed cytoskeletal protein, has been implicated in integrin activation, and its absence is embryonically lethal in mice. In the present study, we tested whether hemostasis might be perturbed in kindlin-2(+/-) mice. Bleeding time and carotid artery occlusion time were significantly prolonged in kindlin-2(+/-) mice. Whereas plasma concentrations/activities of key coagulation/fibrinolytic proteins and platelet counts and aggregation were similar in wild-type and kindlin-2(+/-) mice, kindlin-2(+/-) endothelial cells (ECs) showed enhanced inhibition of platelet aggregation induced by adenosine 5'-diphosphate (ADP) or low concentrations of other agonists. Cell-surface expression of 2 enzymes involved in ADP/adenosine 5'-monophosphate (AMP) degradation, adenosine triphosphate (ATP) diphosphohydrolase (CD39) and ecto-5'-nucleotidase (CD73) were increased twofold to threefold on kindlin-2(+/-) ECs, leading to enhanced ATP/ADP catabolism and production of adenosine, an inhibitor of platelet aggregation. Trafficking of CD39 and CD73 at the EC surface was altered in kindlin-2(+/-) mice. Mechanistically, this was attributed to direct interaction of kindlin-2 with clathrin heavy chain, thereby controlling endocytosis and recycling of CD39 and CD73. The interaction of kindlin-2 with clathrin was independent of its integrin binding site but still dependent on a site within its F3 subdomain. Thus, kindlin-2 regulates trafficking of EC surface enzymes that control platelet responses and hemostasis.
authors.affiliation: Joseph J. Jacobs Center for Thrombosis and Vascular Biology, Department of Molecular Cardiology, and.
authors.forename: Elzbieta
authors.initials: E
authors.lastname: Pluskota
authors.forename: Yi
authors.initials: Y
authors.lastname: Ma
authors.forename: Kamila M
authors.initials: KM
authors.lastname: Bledzka
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Bialkowska
authors.forename: Dmitry A
authors.initials: DA
authors.lastname: Soloviev
authors.forename: Dorota
authors.initials: D
authors.lastname: Szpak
authors.forename: Eugene A
authors.initials: EA
authors.lastname: Podrez
authors.forename: Paul L
authors.initials: PL
authors.lastname: Fox
authors.forename: Stanley L
authors.initials: SL
authors.lastname: Hazen
authors.forename: James J
authors.initials: JJ
authors.lastname: Dowling
authors.forename: Yan-Qing
authors.initials: YQ
authors.lastname: Ma
authors.forename: Edward F
authors.initials: EF
authors.lastname: Plow
mesh_terms: D015720:5'-Nucleotidase / Q000096:biosynthesis; D000244:Adenosine Diphosphate / Q000378:metabolism; D000249:Adenosine Monophosphate / Q000378:metabolism; D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis; D001081:Apyrase / Q000096:biosynthesis; D001792:Blood Platelets / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D002966:Clathrin / Q000378:metabolism*; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D042783:Endothelial Cells / Q000378:metabolism*; D005260:Female; D005434:Flow Cytometry; D006487:Hemostasis / Q000502:physiology*; D047468:Immunoprecipitation; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D009124:Muscle Proteins / Q000378:metabolism*; D010974:Platelet Aggregation / Q000502:physiology; D021381:Protein Transport / Q000502:physiology; D020349:Surface Plasmon Resonance
chemical_list: D015703:Antigens, CD; D002966:Clathrin; D003598:Cytoskeletal Proteins; D009124:Muscle Proteins; C527802:kindlin-2 protein, mouse; D000249:Adenosine Monophosphate; D000244:Adenosine Diphosphate; D015720:5'-Nucleotidase; D001081:Apyrase; C067869:CD39 antigen
references.citation: Nat Med. 1999 Sep;5(9):1010-7
references.pmid: 10470077
references.citation: J Mol Biol. 2010 Nov 26;404(2):274-90
references.pmid: 20875424
references.citation: J Cell Biol. 1993 Dec;123(5):1107-17
references.pmid: 8245121
references.citation: BMC Cell Biol. 2008 Jul 08;9:36
references.pmid: 18611274
references.citation: Biochem Biophys Res Commun. 1992 Mar 16;183(2):803-8
references.pmid: 1550585
references.citation: Blood. 1996 Feb 15;87(4):1238-42
references.pmid: 8608210
references.citation: J Biol Chem. 2012 Jul 13;287(29):24585-94
references.pmid: 22648415
references.citation: Nat Med. 2008 Mar;14(3):325-30
references.pmid: 18278053
references.citation: J Cell Biol. 2003 Feb 3;160(3):321-8
references.pmid: 12551953
references.citation: Science. 2009 May 15;324(5929):895-9
references.pmid: 19443776
references.citation: Am J Transplant. 2010 Dec;10(12):2586-95
references.pmid: 20840479
references.citation: J Clin Invest. 2004 May;113(10):1440-6
references.pmid: 15146241
references.citation: J Biol Chem. 2010 Apr 23;285(17):12559-70
references.pmid: 20189992
references.citation: Curr Opin Hematol. 2009 Sep;16(5):323-8
references.pmid: 19553810
references.citation: EMBO Rep. 2008 Dec;9(12):1203-8
references.pmid: 18997731
references.citation: Trends Cell Biol. 2009 Oct;19(10):504-13
references.pmid: 19766491
references.citation: J Biol Chem. 2005 Jul 15;280(28):26406-14
references.pmid: 15890655
references.citation: Nat Rev Mol Cell Biol. 2011 Jul 22;12(8):517-33
references.pmid: 21779028
references.citation: Circulation. 2005 Feb 15;111(6):808-15
references.pmid: 15699260
references.citation: J Cell Biol. 2008 May 5;181(3):439-46
references.pmid: 18458155
references.citation: J Clin Invest. 1999 Jan;103(2):229-38
references.pmid: 9916135
references.citation: Nat Med. 2009 Mar;15(3):306-12
references.pmid: 19234463
references.citation: Blood. 2010 May 20;115(20):4011-7
references.pmid: 20228270
references.citation: Semin Thromb Hemost. 2005 Apr;31(2):234-46
references.pmid: 15852226
references.citation: Blood. 2011 May 5;117(18):4978-87
references.pmid: 21378273
references.citation: J Exp Med. 2005 Jul 18;202(2):271-81
references.pmid: 16009717
references.citation: Biochem J. 2004 Jan 1;377(Pt 1):1-16
references.pmid: 14505490
references.citation: Nat Med. 2009 Mar;15(3):313-8
references.pmid: 19234460
references.citation: Curr Vasc Pharmacol. 2008 Jan;6(1):52-60
references.pmid: 18220940
references.citation: Nat Rev Mol Cell Biol. 2005 Feb;6(2):112-26
references.pmid: 15687999
references.citation: Cell. 1992 Apr 3;69(1):129-38
references.pmid: 1555237
references.citation: Genes Dev. 2008 May 15;22(10):1325-30
references.pmid: 18483218
references.citation: J Cell Sci. 2011 Mar 15;124(Pt 6):879-91
references.pmid: 21325030
references.citation: Ann N Y Acad Sci. 1983 Jun 27;408:556-66
references.pmid: 6307106
references.citation: J Biol Chem. 2011 Dec 16;286(50):43334-42
references.pmid: 22030399
references.citation: Nat Med. 2007 Sep;13(9):1086-95
references.pmid: 17721545
references.citation: Circ Res. 2004 Oct 15;95(8):814-21
references.pmid: 15358667
references.citation: Proc Natl Acad Sci U S A. 1996 Jun 25;93(13):6258-63
references.pmid: 8692802
references.citation: Circ Res. 2008 Feb 29;102(4):423-31
references.pmid: 18174465
references.citation: Purinergic Signal. 2006 Jun;2(2):351-60
references.pmid: 18404475
references.citation: PLoS Genet. 2008 Dec;4(12):e1000289
references.pmid: 19057668
references.citation: J Cell Biol. 1989 Apr;108(4):1331-43
references.pmid: 2564398
references.citation: J Biol Chem. 2010 Jun 11;285(24):18640-9
references.pmid: 20378539
references.citation: J Clin Invest. 2002 Apr;109(8):1031-40
references.pmid: 11956240
references.citation: Arterioscler Thromb Vasc Biol. 2001 Jul;21(7):1251-2
references.pmid: 11451760
references.citation: J Clin Invest. 1998 May 1;101(9):1851-9
references.pmid: 9576748