G protein coupling of human platelet V1 vascular vasopressin receptors.
G protein coupling of human platelet V1 vascular vasopressin receptors.
复制标题
人血小板 V1 血管加压素受体的 G 蛋白偶联。
DOI：
10.1152/ajpcell.1993.264.5.c1336
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发表时间：
1993
期刊：
影响因子：
--
通讯作者：
Berti-Mattera,L
中科院分区：
文献类型：
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作者：
Thibonnier,M;Goraya,T;Berti-Mattera,L
我们使用了几种方法来确定G蛋白偶联到V1血管精氨酸加压素（AVP）受体的人血小板。在纯化的血小板膜，高亲和力特异性结合的[3 H]AVP，但不是V1血管拮抗剂[3 H]d（CH 2）5 Tyr（Me）AVP的鸟苷5 '-O-（3-硫代三磷酸）或氟化钠调制在MgCl 2的存在和不存在。AVP未能改变[α-32 P]GTP标记模式或24- 27-kDa GTP结合蛋白的胞质易位。AVP刺激的血小板膜GT3活性被Gq α蛋白COOH末端特异性抗体阻断。AVP增加了光反应性GTP类似物[α-32 P]azidoanilido GTP对42 kDa血小板膜蛋白的标记。血小板蛋白与各种G蛋白特异性抗体的免疫印迹显示，42-kDa的蛋白标记的[α-32 P]azidoanilido GTP的免疫印迹仅由抗体特异性的α-亚基的GQ-11。因此，人血小板的V1血管AVP受体以二价阳离子依赖性方式与属于Gq-11家族的G蛋白偶联。
We used several approaches to identify the G protein coupled to V1 vascular arginine vasopressin (AVP) receptors of human platelets. In purified platelet membranes, high-affinity specific binding of [3H]AVP but not that of the V1 vascular antagonist [3H]d(CH2)5Tyr(Me)AVP was modulated by guanosine 5'-O-(3-thiotriphosphate) or sodium fluoride both in the presence and absence of MgCl2. AVP failed to modify the [alpha-32P]GTP labeling pattern or the cytosolic translocation of the 24- to 27-kDa GTP-binding proteins. AVP-stimulated GTPase activity of platelet membranes was blocked by antibodies specific for the COOH-terminal of the Gq alpha protein. AVP increased labeling of a 42-kDa platelet membrane protein by the photoreactive GTP analogue [alpha-32P]azidoanilido GTP. Immunoblotting of platelet proteins with various G protein-specific antibodies revealed that the 42-kDa protein labeled with [alpha-32P]azidoanilido GTP was immunoblotted only by antibodies specific for the alpha-subunit of GQ-11. Thus V1 vascular AVP receptors of human platelets are coupled in a divalent cation-dependent manner to a G protein belonging to the Gq-11 family.