In vitro translation of the human insulin proreceptor results in N-linked glycosylation without dimer formation.
In vitro translation of the human insulin proreceptor results in N-linked glycosylation without dimer formation.
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人胰岛素前受体的体外翻译导致 N-连接糖基化，而不形成二聚体。
DOI：
10.1006/bbrc.1993.1579
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发表时间：
1993
影响因子：
3.1
通讯作者：
Arakaki,RF
中科院分区：
文献类型：
--
作者：
Zhou,X;Baker,NK;Arakaki,RF
细胞表面人胰岛素受体(HINSR)的异四聚体构型对于介导胰岛素的作用非常重要。前受体二聚化是生物发生过程中的第四级加工事件，它的研究为研究二硫键连接的受体亚基之间的相互作用提供了可能性。因此，在利用兔网织红细胞裂解物的转录和翻译耦合方法的无细胞系统中，研究了前受体的二聚体的形成。翻译产物用[35S]蛋氨酸标记，用非还原十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法和放射自显影鉴定。在氧化型谷胱甘肽存在下的体外合成未能证明新生前受体的二聚化。添加微粒体膜的共翻译处理导致前受体的N-连接糖基化，但没有形成二聚体。因此，与在体内观察到的相似，胰岛素前受体二聚化似乎不是共翻译或早期翻译后事件。
The heterotetrameric configuration of the cell surface human insulin receptor (hINSR) is important for mediating insulin action. Investigation of proreceptor dimerization, the quaternary processing event during biogenesis, offers the potential to examine interactions between disulfide-linked receptor subunits. Thus, dimer formation of the proreceptor was examined in a cell-free system that utilized a coupled transcription and translation method with rabbit reticulocyte lysate. Translation products were labeled with [35S]methionine and identified by non-reducing SDS-polyacrylamide gel electrophoresis and autoradiography. In vitro synthesis in the presence of oxidized glutathione failed to demonstrate dimerization of the nascent proreceptor. Co-translational processing with the addition of microsomal membranes resulted in N-linked glycosylation of the proreceptor but without dimer formation. Thus, similar to that observed in vivo, insulin proreceptor dimerization does not appear to be a co-translational or early post-translational event.