Thromb Haemost 2008; 99(01): 96-107
DOI: 10.1160/TH07-08-0498
Platelets and Blood Cells
Schattauer GmbH

Discrete functional motifs reside within the cytoplasmic tail of αV integrin subunit

Thomas A Haas
1   Department of Anatomy and Cell Biology and Molecular Design Research Group, College of Medicine, University of Saskatchewan, Saskatoon, Canada
› Institutsangaben
Financial support: This work was supported by a Canadian Research Chair award, and Canadian Institutes of Health Research and Saskatchewan Health Research Foundation grants.
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Publikationsverlauf

Received: 03. Juli 2007

Accepted after major revision: 07. Oktober 2007

Publikationsdatum:
24. November 2017 (online)

Summary

Previous studies have demonstrated that cell-permeable cytoplasmic tail (CT) αIIβ peptides can modulate the activation of αIIbb3.As αVCT contains an αIIβ homologous region, a series of cell-permeable αV and αIIb peptides were generated to determine if αV CT can modulate the activation of β3 integrins in comparison to αIIb, and to identify the minimal bioactive sequences in αV CT. Using NMR structures and molecular models as guides, the initial peptides for study encompassed the αIIβ homologous sequences of αV CT (αV(987–1006); V-1), its amino-terminus (αV(987–993);V-2), a turn motif (αV(993–1001);V-3), the carboxyl- terminus (αV(999–1006); V-4), and corresponding homologous αIIb peptides. Treatment of platelets and αVβ3-expressing cells with the peptides revealed that IIb-1 inhibited αIIβ3 activation and V-1 inhibited αVβ3 activation, but not vice versa. The inhibitory capacity of these peptides was mapped to the central turn-motif region which was encompassed by V-3, but only partially by IIb-3.V-2 and IIb-2 activated both β3 integrins, while V-4 and IIb-4 were inactive. The use of truncation and mutant peptides confirmed the importance of the turn motif for inhibitory activity and identified the side-chain of αV(Q1001) as a critical inhibitory residue. The difference in the integrin inhibitory capacity of αV and αIIb peptides and their capacity to influence the assembly of kinases with integrin CTs, reveals a possible divergence in the regulatory control of the two β3 integrins.

 
  • References

  • 1 Hynes RO. Integrins: versatility, modulation, and signaling in cell adhesion. Cell 1992; 69: 11-25.
  • 2 Ruoslahti E. Integrins. J Clin Invest 1991; 87: 1-5.
  • 3 Hughes PE, Pfaff M. Integrin affinity modulation. Trends Cell Biol 1998; 8: 359-364.
  • 4 Chen Y-P, Djaffar I, Pidard D. et al. Ser-752 to Pro mutation in the cytoplasmic domain of integrin β3 subunit and defective activation of platelet integrin αIIbβ3 (glycoprotein IIb-IIIa) in a variant of Granzmann thrombasthenia. Proc Natl Acad Sci USA 1992; 89: 10169-10173.
  • 5 O'Toole TE, Katagiri Y, Faull RJ. et al. Integrin cytoplasmic domains mediate inside-out signal transduction. J Cell Biol 1994; 124: 1047-1059.
  • 6 Xiong JP, Stehle T, Diefenbach B. et al. Crystal structure of the extracellular segment of integrin αVβ3 . Science 2001; 294: 339-345.
  • 7 Xiong JP, Stehle T, Zhang R. et al. Crystal structure of the extracellular segment of integrin aV b3 in complex with an Arg-Gly-Asp ligand. Science 2002; 296: 151-155.
  • 8 Xiao T, Takagi J, Coller BS. et al. Structural basis for allostery in integrins and binding to fibrinogen-mimetic therapeutics. Nature 2004; 432: 59-65.
  • 9 Yamanouchi J, Hato T, Tamura T. et al. Identification of critical residues for ligand binding in the integrin β3 I-domain by site-directed mutagenesis. Thromb Haemost 2002; 87: 756-762.
  • 10 Perkins KB, Loftus JC. A mutation in the integrin αIIb subunit that selectively inhibits αIIbβ3 receptor function. Thromb Haemost 2003; 90: 853-862.
  • 11 Tamura T, Hato T, Yamanouchi J. et al. Critical residues for ligand binding in blade 2 of the propeller domain of the integrin αIIb subunit. Thromb Haemost 2004; 91: 111-118.
  • 12 Honda S, Kashiwagi H, Kiyoi T. et al. Amino acid mutagenesis within ligand-binding loops in alpha v confers loss-of-function or gain-of-function phenotype on integrin αvβ3 . Thromb Haemost 2004; 92: 1092-1098.
  • 13 Phillips DR, Nannizzi-Alaimo L, Prasad KS. β3 tyrosine phosphorylation in αIIbβ3 (platelet membrane GP IIb-IIIa) outside-in integrin signaling. Thromb Haemost 2001; 86: 246-258.
  • 14 Shattil SJ, Gao J, Kashiwagi H. Not just another pretty face: regulation of platelet function at the cytoplasmic face of integrin αIIbβ3 . Thromb Haemost 1997; 78: 220-225.
  • 15 Hughes PE, Diaz-Gonzalez F, Leong L. et al. Breaking the integrin hinge: a defined structural constraint regulates integrin signaling. J Biol Chem 1996; 271: 6571-6574.
  • 16 Helluin O, Chan C, Vilaire G. et al. The activation state of αvβ3 regulates platelet and lymphocyte adhesion to intact and thrombin-cleaved osteopontin. J Biol Chem 2000; 275: 18337-18343.
  • 17 Pampori N, Hato T, Stupack DG. et al. Mechanisms and consequences of affinity modulation of integrin aVb3 detected with a novel patch-engineered monovalent ligand. J Biol Chem 1999; 274: 21609-21616.
  • 18 Hughes PE, O’Toole TE, Ylänne J. et al. The conserved membrane-proximal region of an integrin cytoplasmic domain specifies ligand binding affinity. J Biol Chem 1995; 270: 12411-12417.
  • 19 Finberg RW, Cheresh DA. A β turn in the cytoplasmic tail of the integrin αv subunit influences conformation and ligand binding of αvβ3 . J Biol Chem 1994; 269: 4641-4647.
  • 20 Leisner TM, Wencel-Drake JD, Wang W. et al. Bidirectional transmembrane modulation of integrin αIIbβ3 conformations. J Biol Chem 1999; 274: 12945-12949.
  • 21 Haas TA, Plow EF. Development of a structural model for the cytoplasmic domain of an integrin. Protein Eng 1997; 10: 1395-1405.
  • 22 Vallar L, Melchior C, Plancon S. et al. Divalent cations differentially regulate integrin aIIb cytoplasmic tail binding to β3 and to calcium- and integrin-binding protein. J Biol Chem 1999; 274: 17257-17266.
  • 23 Haas TA, Plow EF. The cytoplasmic domain of αIIbβ3: a ternary complex of the integrin α and β subunits and a divalent cation. J Biol Chem 1996; 271: 6017-6026.
  • 24 Vinogradova O, Haas TA, Plow EF. et al. A structural basis for integrin activation by the cytoplasmic tail of the aIIb-subunit. Proc Natl Acad Sci USA 2000; 97: 1450-1455.
  • 25 Vinogradova O, Velyvis A, Velyviene A. et al. A structural mechanism of integrin αIIbβ3 'inside-out' activation as regulated by its cytoplasmic face. Cell 2002; 110: 587-597.
  • 26 Otey CA, Pavalko FM, Burridge K. An interaction between α-actin and the β1 integrin subunit in vitro. J Cell Biol 1990; 111: 721-729.
  • 27 Obergfell A, Eto K, Mocsai A. et al. Coordinate interactions of Csk, Src, and Syk kinases with αlIbβ3 initiate integrin signaling to the cytoskeleton. J Cell Biol 2002; 157: 265-275.
  • 28 Calderwood DA, Zent R, Grant R. et al. The talin head domain binds to integrin b subunit cytoplasmic tails and regulates integrin activation. J Biol Chem 1999; 274: 28071-28074.
  • 29 Shattil SJ, O'Toole TE, Eigenthaler M. et al. β3-Endonexin, a novel polypeptide that interacts specifically with the cytoplasmic tail of the integrin β3 subunit. J Cell Biol 1995; 131: 807-816.
  • 30 Reddy KB, Gascard P, Price MG. et al. Identification of an interaction between the M-band protein Skelemin and beta -integrin subunits. Colocalization of a Skelemin-like protein with beta 1– and beta 3-integrins in non-muscle cells. J Biol Chem 1998; 273: 35039-35047.
  • 31 Kieffer JD, Plopper G, Ingber DE. et al. Direct binding of F actin to the cytoplasmic domain of the a2 integrin chain in vitro. Biochem Biophys Res Commun 1995; 217: 466-474.
  • 32 Knezevic I, Leisner TM, Lam SCT. Direct binding of the platelet integrin αIIbβ3 (GPIIb- IIIa) to talin. Evidence that interaction is mediated through the cytoplasmic domains of both αIIb and β3. Biol Chem 1996; 271: 16416-16421.
  • 33 Naik UP, Patel PM, Parise LV. Identification of a novel calcium-binding protein that interacts with the integrin aIIb cytoplasmic domain. J Biol Chem 1997; 272: 4651-4654.
  • 34 Kato A, Kawamata N, Tamayose K. et al. Ancient ubiquitous protein 1 binds to the conserved membraneproximal sequence of the cytoplasmic tail of the integrin α subunits that plays a crucial role in the inside-out signaling of αIIbβ3 . J Biol Chem 2002; 277: 28934-28941.
  • 35 Larkin D, Murphy D, Reilly DF. et al. ICln, a novel integrin αIIbβ3-associated protein, functionally regulates platelet activation. J Biol Chem 2004; 279: 27286-27293.
  • 36 Vijayan KV, Liu Y, Li TT. et al. Protein phosphatase 1 associates with the integrin aIIb subunit and regulates signaling. J Biol Chem 2004; 279: 33039-33042.
  • 37 Yuan W, Leisner TM, McFadden AW. et al. CIB1 is an endogenous inhibitor of agonist-induced integrin αIIbb3 activation. J Cell Biol 2006; 172: 169-175.
  • 38 Stephens G, O’Luanaigh N, Reilly D. et al. A sequence within the cytoplasmic tail of αIIb independently activates platelet aggregation and thromboxane synthesis. J Biol Chem 1998; 273: 20317-20322.
  • 39 Aylward K, Meade G, Ahrens I. et al. A novel functional role for the highly conserved a-subunit KVGFFKR motif distinct from integrin αIIbβ3 activation processes. J Thromb Haemost 2006; 4: 1804-12-1812.
  • 40 Barry WT, Boudignon-Proudhon C, Shock DD. et al. Molecular basis of CIB binding to the integrin αIIb cytoplasmic domain. J Biol Chem 2002; 277: 28877-28883.
  • 41 Ginsberg MH, Yaspan B, Forsyth J. et al. A membrane- distal segment of the integrin aIIb cytoplasmic domain regulates integrin activation. J Biol Chem 2001; 276: 22514-22521.
  • 42 Plow EF, Srouji AH, Meyer D. et al. Evidence that three adhesive proteins interact with common recognition site on activated platelets. J Biol Chem 1984; 259: 5388-5391.
  • 43 Yan B, Calderwood DA, Yaspan B. et al. Calpain cleavage promotes talin binding to the b3 integrin cytoplasmic domain. J Biol Chem 2001; 276: 28164-28170.
  • 44 D'Souza SE, Haas TA, Piotrowicz RS. et al. Ligand and cation binding are dual functions of a discrete segment of the integrin b3 subunit: cation displacement is involved in ligand binding. Cell 1994; 79: 659-667.
  • 45 Porte F, Liautard J.-P, Köhler S. Early acidification of phagosomes containing Brucella suis is essential for intracellular survival in murine macrophages. Infect Immun 1999; 67: 4041-4047.
  • 46 Patil S, Jedsadayanmata A, Wencel-Drake JD. et al. Identification of a talin-binding site in the integrin b3 subunit distinct from the NPLY regulatory motif of post-ligand binding functions. The talin n-terminal head domain interacts with the membrane-proximal region of the b3 cytoplasmic tail. J Biol Chem 1999; 274: 28575-28583.
  • 47 Schlaepfer DD, Hunter T. Focal Adhesion Kinase Overexpression Enhances Ras-dependent Integrin Signaling to ERK2/Mitogen-activated Protein Kinase through Interactions with and Activation of c-Src. J Biol Chem 1997; 272: 13189-13195.