ADAP interactions with talin and kindlin promote platelet integrin alpha IIb beta 3 activation and stable fibrinogen binding

作者：Kasirer Friede Ana^*; Kang Jian; Kahner Bryan; Ye Feng; Gin**erg Mark H; Shattil Sanford J

来源：Blood, 2014, 123(20): 3156-3165.

DOI：10.1182/blood-2013-08-520627

ADAP is a hematopoietic-restricted adapter protein that promotes integrin activation and is a carrier for other adapter proteins, Src kinase-associated phosphoprotein 1 (SKAP1) and SKAP2. In T lymphocytes, SKAP1 is the ADAP-associated molecule that activates integrins through direct linkages with Rap1 effectors (regulator of cell adhesion and polarization enriched in lymphoid tissues; Rap1-interacting adapter molecule). ADAP also promotes integrin alpha IIb beta 3 activation in platelets, which lack SKAP1, suggesting an ADAP integrin-regulatory pathway different from those in lymphocytes. Here we characterized a novel association between ADAP and 2 essential integrin-beta cytoplasmic tail-binding proteins involved in alpha IIb beta 3 activation, talin and kindlin-3. Glutathione S-transferase pull-downs identified distinct regions in ADAP necessary for association with kindlin or talin. ADAP was physically proximal to talin and kindlin-3 in human platelets, as assessed biochemically, and by immunofluorescence microscopy and proximity ligation. Relative to wild-type mouse platelets, ADAP-deficient platelets exhibited reduced co-localization of talin with alpha IIb beta 3, and reduced irreversible fibrinogen binding in response to a protease activated receptor 4 (PAR4) thrombin receptor agonist. When ADAP was heterologously expressed in Chinese hamster ovary cells co-expressing alpha IIb beta 3, talin, PAR1, and kindlin-3, it associated with an alpha IIb beta 3/talin complex and enabled kindlin-3 to promote agonist-dependent ligand binding to alpha IIb beta 3. Thus, ADAP uniquely promotes activation of and irreversible fibrinogen binding to platelet alpha IIb beta 3 through interactions with talin and kindlin-3.

• 出版日期2014-5-15