McClue S J, Milligan G
Department of Biochemistry, University of Glasgow, Scotland, UK.
FEBS Lett. 1990 Sep 3;269(2):430-4. doi: 10.1016/0014-5793(90)81209-7.
In membranes of undifferentiated neuroblastoma x glioma hybrid cell line NG108-15, the apparent specific binding of [3H]yohimbine measured in the presence of 1 microM noradrenaline, was increased substantially by the presence of the poorly hydrolysed analogue of GTP, guanylyl-imidodiphosphate (Gpp[NH]p) or by preincubation of membranes with antibodies against the C-terminal decapeptide of the alpha subunit of the G-protein Gi2. Such an effect was not produced by antibodies against the equivalent region of Go alpha Gi3 alpha or Gs alpha or from non-immune serum. By contrast, total specific binding of [3H]yohimbine was not modified by co-incubation with Gpp[NH]p or by preincubation with the antibodies from any of the anti-G protein antisera. These results demonstrate a direct interaction of the alpha 2B adrenergic receptor of NG108-15 cells with Gi2.
在未分化的神经母细胞瘤x胶质瘤杂交细胞系NG108 - 15的细胞膜中,在1微摩尔去甲肾上腺素存在的情况下测得的[3H]育亨宾的表观特异性结合,在存在水解能力差的GTP类似物鸟苷酰-亚氨基二磷酸(Gpp[NH]p)时,或通过用针对G蛋白Gi2α亚基C末端十肽的抗体对细胞膜进行预孵育后,会显著增加。针对Goα、Gi3α或Gsα的等效区域的抗体或非免疫血清不会产生这种效应。相比之下,[3H]育亨宾的总特异性结合不会因与Gpp[NH]p共同孵育或用任何抗G蛋白抗血清的抗体进行预孵育而改变。这些结果证明了NG108 - 15细胞的α2B肾上腺素能受体与Gi2之间存在直接相互作用。