Van der Ploeg I, Fredholm B B, Carlquist M, Bertorello A, Aperia A, Celsi G
Department of Pharmacology, Karolinska Institutet, Stockholm, Sweden.
Acta Physiol Scand. 1991 May;142(1):83-90. doi: 10.1111/j.1748-1716.1991.tb09131.x.
Dopamine was shown to affect Na+,K(+)-ATPase activity in basolateral membranes of the rat kidney via a pertussis toxin dependent mechanism. In order to examine if some form of pertussis toxin sensitive G-protein is present exclusively in the basolateral membrane of the rat renal cortex we examined the G-protein composition of both apical and basolateral membrane vesicles. Western blots showed an essentially uniform distribution of G alpha total, G alpha S and G beta over the two membranes. Go could not be detected with western blot technique in the vesicle preparations. By contrast, the distribution of ADP-ribosylation with the bacterial toxins pertussis toxin and cholera toxin depended on the amount of detergent in the assay and perhaps other factors, and thus could not be used to evaluate the relative amounts of G-protein subunits. Thus, in contrast to the situation in cultured renal cells, unequal distribution of receptor and G-protein substrates is apparently not paralleled by an unequal distribution of the detected forms of G-proteins under physiological conditions.
多巴胺通过一种百日咳毒素依赖性机制影响大鼠肾脏基底外侧膜中的Na +,K(+)-ATP酶活性。为了研究某种形式的百日咳毒素敏感G蛋白是否仅存在于大鼠肾皮质的基底外侧膜中,我们检测了顶端和基底外侧膜囊泡的G蛋白组成。蛋白质免疫印迹显示Gα总量、GαS和Gβ在两种膜上基本均匀分布。在囊泡制剂中,用蛋白质免疫印迹技术无法检测到Go。相比之下,细菌毒素百日咳毒素和霍乱毒素的ADP核糖基化分布取决于测定中去污剂的量以及其他因素,因此不能用于评估G蛋白亚基的相对含量。因此,与培养的肾细胞情况不同,在生理条件下,受体和G蛋白底物的不均匀分布显然与检测到的G蛋白形式的不均匀分布不平行。