THE IDENTIFICATION AND LOCALIZATION OF THE GUANINE-NUCLEOTIDE BINDING-PROTEIN G0 IN THE AUDITORY-SYSTEM
THE IDENTIFICATION AND LOCALIZATION OF THE GUANINE-NUCLEOTIDE BINDING-PROTEIN G0 IN THE AUDITORY-SYSTEM
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DOI：
10.1111/j.1460-9568.1991.tb00066.x
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发表时间：
1991-12-01
影响因子：
3.4
通讯作者：
BOCKAERT, J
中科院分区：
文献类型：
--
作者：
CANLON, B;HOMBURGER, V;BOCKAERT, J
豚鼠组织中鸟嘌呤核苷酸结合蛋白（G蛋白）的鉴定通过百日咳博德特氏菌毒素对α亚基的腺苷二磷酸核糖基化进行评估，使用[α-P-32]烟酰胺腺嘌呤二核苷酸作为底物，然后进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和放射自显影。 三种组织（下丘、神经母细胞瘤细胞和柯蒂氏器）含有 G0-α (39 kD)、Gi2-α (40 kD) 和 Gi1-α 和/或 Gi3-α (41 kD)。 血管纹和第八神经主要含有Gi2-α、Gi1-α和/或Gi3-α，但几乎检测不到G0-α。 来自柯蒂氏器的外毛细胞纯化制剂含有所有三种百日咳毒素底物，包括 G0-α，其中最突出的是 Gi2-α (40 kD) 亚基。 将分离的外毛细胞、Hensen 细胞和血管纹与亲和纯化的抗 G0-α 抗体一起孵育后，通过光学显微镜确定 G0-α 亚基的免疫细胞化学定位。 在毛细胞中，沿着质膜和角质板（粘附带）周边观察到阳性反应。 在从角质板延伸至外毛细胞核的角质层网络内也观察到阳性反应。 最后，外毛细胞的基部还含有 G0-α。 然而，该细胞区域中存在的 G0-α 很可能不在毛细胞本身内，而是在解剖过程中保持附着的神经末梢中。
The identification of guanine nucleotide binding proteins (G proteins) in guinea-pig tissues was assessed by the adenosine diphosphate-ribosylation of the alpha-subunit by Bordetella pertussis toxin using [alpha-P-32]nicotinamide adenine dinucleotide as the substrate followed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and autoradiography. Three tissues (inferior colliculus, neuroblastoma cells, and the organ of Corti) contained G0-alpha (39 kD), as well as Gi2-alpha (40 kD) and Gi1-alpha and/or Gi3-alpha (41 kD). The stria vascularis and the VIIIth nerve contained mainly Gi2-alpha, Gi1-alpha and/or Gi3-alpha, but G0-alpha was barely detectable. A purified preparation of outer hair cells from the organ of Corti contained all three pertussis toxin substrates including G0-alpha, with the Gi2-alpha (40 kD) subunit being the most prominent. The immunocytochemical localization of the G0-alpha subunit was determined by light microscopy after incubating isolated outer hair cells, Hensen cells and the stria vascularis with affinity-purified anti-G0-alpha antibodies. In hair cells a positive reaction was observed along the plasma membrane and around the perimeter of the cuticular plate (zona adherens). Positive reaction was also observed within the infracuticular network extending from the cuticular plate towards the nucleus in outer hair cells. Finally, the base of the outer hair cells also contained G0-alpha. However, it is likely that the G0-alpha that is present in this cell region is not within the hair cell itself, but rather in nerve terminals which remained attached during dissection.