Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 2004 Apr 15;379(Pt 2):317–323. doi: 10.1042/BJ20031753

Suppression of integrin activation by the membrane-distal sequence of the integrin alphaIIb cytoplasmic tail.

Jun Yamanouchi 1, Takaaki Hato 1, Tatsushiro Tamura 1, Shigeru Fujita 1
PMCID: PMC1224082  PMID: 14723599

Abstract

Integrin cytoplasmic tails regulate integrin activation including an increase in integrin affinity for ligands. Although there is ample evidence that the membrane-proximal regions of the alpha and beta tails interact with each other to maintain integrins in a low-affinity state, little is known about the role of the membrane-distal region of the alpha tail in regulation of integrin activation. We report a critical sequence for regulation of integrin activation in the membrane-distal region of the alphaIIb tail. Alanine substitution of the RPP residues in the alphaIIb tail rendered alphaIIbbeta3 constitutively active in a metabolic energy-dependent manner. Although an alphaIIb/alpha6Abeta3 chimaeric integrin, in which the alphaIIb tail was replaced by the alpha6A tail, was in an energy-dependent active state to bind soluble ligands, introduction of the RPP sequence into the alpha6A tail inhibited binding of an activation-dependent antibody PAC1. In alphaIIb/alpha6Abeta3, deleting the TSDA sequence from the alpha6A tail or single amino acid substitutions of the TSDA residues inhibited alphaIIb/alpha6Abeta3 activation and replacing the membrane-distal region of the alphaIIb tail with TSDA rendered alphaIIbbeta3 active, suggesting a stimulatory role of TSDA in energy-dependent integrin activation. However, adding TSDA to the alphaIIb tail containing the RPP sequence of the membrane-distal region failed to activate alphaIIbbeta3. These results suggest that the RPP sequence after the GFFKR motif of the alphaIIb tail suppresses energy-dependent alphaIIbbeta3 activation. These findings provide a molecular basis for the regulation of energy-dependent integrin activation by alpha subunit tails.

Full Text

The Full Text of this article is available as a PDF (141.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abrams C., Deng Y. J., Steiner B., O'Toole T., Shattil S. J. Determinants of specificity of a baculovirus-expressed antibody Fab fragment that binds selectively to the activated form of integrin alpha IIb beta 3. J Biol Chem. 1994 Jul 22;269(29):18781–18788. [PubMed] [Google Scholar]
  2. Barry William T., Boudignon-Proudhon Christel, Shock David D., McFadden Andrew, Weiss Jonathan M., Sondek John, Parise Leslie V. Molecular basis of CIB binding to the integrin alpha IIb cytoplasmic domain. J Biol Chem. 2002 May 21;277(32):28877–28883. doi: 10.1074/jbc.M202983200. [DOI] [PubMed] [Google Scholar]
  3. Bennett J. S., Shattil S. J., Power J. W., Gartner T. K. Interaction of fibrinogen with its platelet receptor. Differential effects of alpha and gamma chain fibrinogen peptides on the glycoprotein IIb-IIIa complex. J Biol Chem. 1988 Sep 15;263(26):12948–12953. [PubMed] [Google Scholar]
  4. Bunn R. C., Jensen M. A., Reed B. C. Protein interactions with the glucose transporter binding protein GLUT1CBP that provide a link between GLUT1 and the cytoskeleton. Mol Biol Cell. 1999 Apr;10(4):819–832. doi: 10.1091/mbc.10.4.819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chen Y. P., O'Toole T. E., Shipley T., Forsyth J., LaFlamme S. E., Yamada K. M., Shattil S. J., Ginsberg M. H. "Inside-out" signal transduction inhibited by isolated integrin cytoplasmic domains. J Biol Chem. 1994 Jul 15;269(28):18307–18310. [PubMed] [Google Scholar]
  6. El Mourabit Haquima, Poinat Patrice, Koster Jan, Sondermann Holger, Wixler Viktor, Wegener Elmar, Laplantine Emmanuel, Geerts Dirk, Georges-Labouesse Elisabeth, Sonnenberg Arnoud. The PDZ domain of TIP-2/GIPC interacts with the C-terminus of the integrin alpha5 and alpha6 subunits. Matrix Biol. 2002 Mar;21(2):207–214. doi: 10.1016/s0945-053x(01)00198-6. [DOI] [PubMed] [Google Scholar]
  7. Fenczik C. A., Sethi T., Ramos J. W., Hughes P. E., Ginsberg M. H. Complementation of dominant suppression implicates CD98 in integrin activation. Nature. 1997 Nov 6;390(6655):81–85. doi: 10.1038/36349. [DOI] [PubMed] [Google Scholar]
  8. García-Alvarez Begoña, de Pereda José M., Calderwood David A., Ulmer Tobias S., Critchley David, Campbell Iain D., Ginsberg Mark H., Liddington Robert C. Structural determinants of integrin recognition by talin. Mol Cell. 2003 Jan;11(1):49–58. doi: 10.1016/s1097-2765(02)00823-7. [DOI] [PubMed] [Google Scholar]
  9. Giancotti F. G., Ruoslahti E. Integrin signaling. Science. 1999 Aug 13;285(5430):1028–1032. doi: 10.1126/science.285.5430.1028. [DOI] [PubMed] [Google Scholar]
  10. Ginsberg M. H., Yaspan B., Forsyth J., Ulmer T. S., Campbell I. D., Slepak M. A membrane-distal segment of the integrin alpha IIb cytoplasmic domain regulates integrin activation. J Biol Chem. 2001 Apr 13;276(25):22514–22521. doi: 10.1074/jbc.M101915200. [DOI] [PubMed] [Google Scholar]
  11. Harris B. Z., Lim W. A. Mechanism and role of PDZ domains in signaling complex assembly. J Cell Sci. 2001 Sep;114(Pt 18):3219–3231. doi: 10.1242/jcs.114.18.3219. [DOI] [PubMed] [Google Scholar]
  12. Hato T., Pampori N., Shattil S. J. Complementary roles for receptor clustering and conformational change in the adhesive and signaling functions of integrin alphaIIb beta3. J Cell Biol. 1998 Jun 29;141(7):1685–1695. doi: 10.1083/jcb.141.7.1685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hughes P. E., Diaz-Gonzalez F., Leong L., Wu C., McDonald J. A., Shattil S. J., Ginsberg M. H. Breaking the integrin hinge. A defined structural constraint regulates integrin signaling. J Biol Chem. 1996 Mar 22;271(12):6571–6574. doi: 10.1074/jbc.271.12.6571. [DOI] [PubMed] [Google Scholar]
  14. Hughes P. E., Pfaff M. Integrin affinity modulation. Trends Cell Biol. 1998 Sep;8(9):359–364. doi: 10.1016/s0962-8924(98)01339-7. [DOI] [PubMed] [Google Scholar]
  15. Hughes P. E., Renshaw M. W., Pfaff M., Forsyth J., Keivens V. M., Schwartz M. A., Ginsberg M. H. Suppression of integrin activation: a novel function of a Ras/Raf-initiated MAP kinase pathway. Cell. 1997 Feb 21;88(4):521–530. doi: 10.1016/s0092-8674(00)81892-9. [DOI] [PubMed] [Google Scholar]
  16. Hynes Richard O. Integrins: bidirectional, allosteric signaling machines. Cell. 2002 Sep 20;110(6):673–687. doi: 10.1016/s0092-8674(02)00971-6. [DOI] [PubMed] [Google Scholar]
  17. Kato Atsushi, Kawamata Norihiko, Tamayose Kenji, Egashira Motoki, Miura Rika, Fujimura Tsutomu, Murayama Kimie, Oshimi Kazuo. Ancient ubiquitous protein 1 binds to the conserved membrane-proximal sequence of the cytoplasmic tail of the integrin alpha subunits that plays a crucial role in the inside-out signaling of alpha IIbbeta 3. J Biol Chem. 2002 May 31;277(32):28934–28941. doi: 10.1074/jbc.M204340200. [DOI] [PubMed] [Google Scholar]
  18. Leisner T. M., Wencel-Drake J. D., Wang W., Lam S. C. Bidirectional transmembrane modulation of integrin alphaIIbbeta3 conformations. J Biol Chem. 1999 Apr 30;274(18):12945–12949. doi: 10.1074/jbc.274.18.12945. [DOI] [PubMed] [Google Scholar]
  19. Li R., Babu C. R., Lear J. D., Wand A. J., Bennett J. S., DeGrado W. F. Oligomerization of the integrin alphaIIbbeta3: roles of the transmembrane and cytoplasmic domains. Proc Natl Acad Sci U S A. 2001 Oct 16;98(22):12462–12467. doi: 10.1073/pnas.221463098. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Liddington R. C., Ginsberg M. H. Integrin activation takes shape. J Cell Biol. 2002 Sep 3;158(5):833–839. doi: 10.1083/jcb.200206011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Liu S., Calderwood D. A., Ginsberg M. H. Integrin cytoplasmic domain-binding proteins. J Cell Sci. 2000 Oct;113(Pt 20):3563–3571. doi: 10.1242/jcs.113.20.3563. [DOI] [PubMed] [Google Scholar]
  22. Lu C., Takagi J., Springer T. A. Association of the membrane proximal regions of the alpha and beta subunit cytoplasmic domains constrains an integrin in the inactive state. J Biol Chem. 2001 Jan 30;276(18):14642–14648. doi: 10.1074/jbc.M100600200. [DOI] [PubMed] [Google Scholar]
  23. O'Toole T. E., Katagiri Y., Faull R. J., Peter K., Tamura R., Quaranta V., Loftus J. C., Shattil S. J., Ginsberg M. H. Integrin cytoplasmic domains mediate inside-out signal transduction. J Cell Biol. 1994 Mar;124(6):1047–1059. doi: 10.1083/jcb.124.6.1047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. O'Toole T. E., Mandelman D., Forsyth J., Shattil S. J., Plow E. F., Ginsberg M. H. Modulation of the affinity of integrin alpha IIb beta 3 (GPIIb-IIIa) by the cytoplasmic domain of alpha IIb. Science. 1991 Nov 8;254(5033):845–847. doi: 10.1126/science.1948065. [DOI] [PubMed] [Google Scholar]
  25. O'Toole T. E., Ylanne J., Culley B. M. Regulation of integrin affinity states through an NPXY motif in the beta subunit cytoplasmic domain. J Biol Chem. 1995 Apr 14;270(15):8553–8558. doi: 10.1074/jbc.270.15.8553. [DOI] [PubMed] [Google Scholar]
  26. Phillips D. R., Nannizzi-Alaimo L., Prasad K. S. Beta3 tyrosine phosphorylation in alphaIIbbeta3 (platelet membrane GP IIb-IIIa) outside-in integrin signaling. Thromb Haemost. 2001 Jul;86(1):246–258. [PubMed] [Google Scholar]
  27. Plow E. F., Haas T. A., Zhang L., Loftus J., Smith J. W. Ligand binding to integrins. J Biol Chem. 2000 Jul 21;275(29):21785–21788. doi: 10.1074/jbc.R000003200. [DOI] [PubMed] [Google Scholar]
  28. Ramos J. W., Kojima T. K., Hughes P. E., Fenczik C. A., Ginsberg M. H. The death effector domain of PEA-15 is involved in its regulation of integrin activation. J Biol Chem. 1998 Dec 18;273(51):33897–33900. doi: 10.1074/jbc.273.51.33897. [DOI] [PubMed] [Google Scholar]
  29. Shattil S. J., Ginsberg M. H. Perspectives series: cell adhesion in vascular biology. Integrin signaling in vascular biology. J Clin Invest. 1997 Jul 1;100(1):1–5. doi: 10.1172/JCI119500. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Shattil S. J., Hoxie J. A., Cunningham M., Brass L. F. Changes in the platelet membrane glycoprotein IIb.IIIa complex during platelet activation. J Biol Chem. 1985 Sep 15;260(20):11107–11114. [PubMed] [Google Scholar]
  31. Shattil S. J., Kashiwagi H., Pampori N. Integrin signaling: the platelet paradigm. Blood. 1998 Apr 15;91(8):2645–2657. [PubMed] [Google Scholar]
  32. Shimaoka Motomu, Xiao Tsan, Liu Jin-Huan, Yang Yuting, Dong Yicheng, Jun Chang-Duk, McCormack Alison, Zhang Rongguang, Joachimiak Andrzej, Takagi Junichi. Structures of the alpha L I domain and its complex with ICAM-1 reveal a shape-shifting pathway for integrin regulation. Cell. 2003 Jan 10;112(1):99–111. doi: 10.1016/s0092-8674(02)01257-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Stephens G., O'Luanaigh N., Reilly D., Harriott P., Walker B., Fitzgerald D., Moran N. A sequence within the cytoplasmic tail of GpIIb independently activates platelet aggregation and thromboxane synthesis. J Biol Chem. 1998 Aug 7;273(32):20317–20322. doi: 10.1074/jbc.273.32.20317. [DOI] [PubMed] [Google Scholar]
  34. Tadokoro Seiji, Shattil Sanford J., Eto Koji, Tai Vera, Liddington Robert C., de Pereda Jose M., Ginsberg Mark H., Calderwood David A. Talin binding to integrin beta tails: a final common step in integrin activation. Science. 2003 Oct 3;302(5642):103–106. doi: 10.1126/science.1086652. [DOI] [PubMed] [Google Scholar]
  35. Takagi J., Erickson H. P., Springer T. A. C-terminal opening mimics 'inside-out' activation of integrin alpha5beta1. Nat Struct Biol. 2001 May;8(5):412–416. doi: 10.1038/87569. [DOI] [PubMed] [Google Scholar]
  36. Tani T. T., Mercurio A. M. PDZ interaction sites in integrin alpha subunits. T14853, TIP/GIPC binds to a type I recognition sequence in alpha 6A/alpha 5 and a novel sequence in alpha 6B. J Biol Chem. 2001 Jul 30;276(39):36535–36542. doi: 10.1074/jbc.M105785200. [DOI] [PubMed] [Google Scholar]
  37. Tokuhira M., Handa M., Kamata T., Oda A., Katayama M., Tomiyama Y., Murata M., Kawai Y., Watanabe K., Ikeda Y. A novel regulatory epitope defined by a murine monoclonal antibody to the platelet GPIIb-IIIa complex (alpha IIb beta 3 integrin). Thromb Haemost. 1996 Dec;76(6):1038–1046. [PubMed] [Google Scholar]
  38. Tsuboi Shigeru. Calcium integrin-binding protein activates platelet integrin alpha IIbbeta 3. J Biol Chem. 2001 Nov 9;277(3):1919–1923. doi: 10.1074/jbc.M110643200. [DOI] [PubMed] [Google Scholar]
  39. Ulmer T. S., Yaspan B., Ginsberg M. H., Campbell I. D. NMR analysis of structure and dynamics of the cytosolic tails of integrin alpha IIb beta 3 in aqueous solution. Biochemistry. 2001 Jun 26;40(25):7498–7508. doi: 10.1021/bi010338l. [DOI] [PubMed] [Google Scholar]
  40. Vinogradova O., Haas T., Plow E. F., Qin J. A structural basis for integrin activation by the cytoplasmic tail of the alpha IIb-subunit. Proc Natl Acad Sci U S A. 2000 Feb 15;97(4):1450–1455. doi: 10.1073/pnas.040548197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Vinogradova Olga, Velyvis Algirdas, Velyviene Asta, Hu Bin, Haas Thomas, Plow Edward, Qin Jun. A structural mechanism of integrin alpha(IIb)beta(3) "inside-out" activation as regulated by its cytoplasmic face. Cell. 2002 Sep 6;110(5):587–597. doi: 10.1016/s0092-8674(02)00906-6. [DOI] [PubMed] [Google Scholar]
  42. Weljie Aalim M., Hwang Peter M., Vogel Hans J. Solution structures of the cytoplasmic tail complex from platelet integrin alpha IIb- and beta 3-subunits. Proc Natl Acad Sci U S A. 2002 Apr 30;99(9):5878–5883. doi: 10.1073/pnas.092515799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Woodside D. G., Liu S., Ginsberg M. H. Integrin activation. Thromb Haemost. 2001 Jul;86(1):316–323. [PubMed] [Google Scholar]
  44. Yamanouchi Jun, Hato Takaaki, Tamura Tatsushiro, Fujita Shigeru. Identification of critical residues for ligand binding in the integrin beta3 I-domain by site-directed mutagenesis. Thromb Haemost. 2002 Apr;87(4):756–762. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES