Analysis of the roles of 14-3-3 in the platelet glycoprotein Ib-IX-mediated activation of integrin αIIbβ3 using a reconstituted mammalian cell expression model

被引:107
作者
Gu, MY
Xi, XD
Englund, GD
Berndt, MC
Du, XP
机构
[1] Univ Illinois, Coll Med, Dept Pharmacol, Chicago, IL 60612 USA
[2] Baker Med Res Inst, Prahran, Vic 3181, Australia
关键词
platelet; glycoprotein Ib-IX; integrin; 14-3-3; von Willebrand factor;
D O I
10.1083/jcb.147.5.1085
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
We have reconstituted the platelet glycoprotein (GP) Ib-IX-mediated activation of the integrin alpha(IIb)beta(3) in a recombinant DNA expression model, and show that 14-3-3 is important in GPIb-IX signaling. CHO cells expressing alpha(IIb)beta(3) adhere poorly to vWF. Cells expressing GPIb-IX adhere to vWF in the presence of botrocetin but spread poorly. Cells coexpressing integrin (alpha(IIb)beta(3) and GPIb-IX adhere and spread on VWF, which is inhibited by RGDS peptides and antibodies against alpha(IIb)beta(3) vWF binding to GPIb-IX also activates soluble fibrinogen binding to alpha(IIb)beta(3) indicating that GPIb-IX mediates a cellular signal leading to alpha(IIb)beta(3) activation. Deletion of the 14-3-3-binding site in GPIb alpha inhibited GPIb-IX-mediated fibrinogen binding to alpha(IIb)beta(3) and cell spreading on vWF. Thus, 14-3-3 binding to GPIb-IX is important in GPIb-IX signaling. Expression of a dominant negative 14-3-3 mutant inhibited cell spreading on vWF, suggesting an important role for 14-3-3. Deleting both the 14-3-3 and filamin-binding sites of GPIb alpha induced an endogenous integrin-dependent cell spreading on VWF without requiring alpha(IIb)beta(3), but inhibited vWF-induced fibrinogen binding to alpha(IIb)beta(3). Thus, while different activation mechanisms may be responsible for vWF interaction with different integrins, GPIb-IX-mediated activation of alpha(IIb)beta(3) requires 14-3-3 interaction with GPIb alpha.
引用
收藏
页码:1085 / 1096
页数:12
相关论文
共 69 条
  • [41] Activation-modulated association of 14-3-3 proteins with Cbl in T cells
    Liu, YC
    Elly, C
    Yoshida, H
    BonnefoyBerard, N
    Altman, A
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (24) : 14591 - 14595
  • [42] Bernard-Soulier syndrome
    López, JA
    Andrews, RK
    Afshar-Kharghan, V
    Berndt, MC
    [J]. BLOOD, 1998, 91 (12) : 4397 - 4418
  • [43] LOPEZ JA, 1994, BLOOD COAGUL FIBRIN, V5, P97
  • [44] Identification of domains responsible for von Willebrand factor type VI collagen interaction mediating platelet adhesion under high flow
    Mazzucato, M
    Spessotto, P
    Masotti, A
    De Appollonia, L
    Cozzi, MR
    Yoshioka, A
    Perris, R
    Colombatti, A
    De Marco, L
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (05) : 3033 - 3041
  • [45] Meller N, 1996, MOL CELL BIOL, V16, P5782
  • [46] Interaction of 14-3-3 with signaling proteins is mediated by the recognition of phosphoserine
    Muslin, AJ
    Tanner, JW
    Allen, PM
    Shaw, AS
    [J]. CELL, 1996, 84 (06) : 889 - 897
  • [47] AFFINITY MODULATION OF THE ALPHA-IIB-BETA-3 INTEGRIN (PLATELET GPIIB-IIIA) IS AN INTRINSIC PROPERTY OF THE RECEPTOR
    OTOOLE, TE
    LOFTUS, JC
    DU, XP
    GLASS, AA
    RUGGERI, ZM
    SHATTIL, SJ
    PLOW, EF
    GINSBERG, MH
    [J]. CELL REGULATION, 1990, 1 (12): : 883 - 893
  • [48] ASSOCIATION OF POLYOMAVIRUS MIDDLE TUMOR-ANTIGEN WITH 14-3-3-PROTEINS
    PALLAS, DC
    FU, H
    HAEHNEL, LC
    WELLER, W
    COLLIER, RJ
    ROBERTS, TM
    [J]. SCIENCE, 1994, 265 (5171) : 535 - 537
  • [49] GPIIB-IIIA - THE RESPONSIVE INTEGRIN
    PHILLIPS, DR
    CHARO, IF
    SCARBOROUGH, RM
    [J]. CELL, 1991, 65 (03) : 359 - 362
  • [50] SHEAR-STRESS-INDUCED VON-WILLEBRAND-FACTOR BINDING TO PLATELETS CAUSES THE ACTIVATION OF TYROSINE KINASE(S)
    RAZDAN, K
    HELLUMS, JD
    KROLL, MH
    [J]. BIOCHEMICAL JOURNAL, 1994, 302 : 681 - 686