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一种在特定神经元中差异表达的Gq类软体动物G蛋白α亚基的克隆。

Cloning of a molluscan G protein alpha subunit of the Gq class which is expressed differentially in identified neurons.

作者信息

Knol J C, Ramnatsingh S, van Kesteren E R, van Minnen J, Planta R J, van Heerikhuizen H, Vreugdenhil E

机构信息

Graduate School of Neurosciences Amsterdam, Research Institute Neurosciences Vrije Universiteit, Faculty of Chemistry, The Netherlands.

出版信息

Eur J Biochem. 1995 May 15;230(1):193-9.

PMID:7601100
Abstract

Through molecular cloning we have identified a molluscan G protein alpha subunit which belongs to the G alpha q family and is expressed in the central nervous system (CNS) of the pond snail, Lymnaea stagnalis. The deduced protein product shares a very high degree of amino sequence identity with vertebrate and invertebrate G alpha q/G alpha 11 subunits (80-82% and 76-77%, respectively). Large parts of the protein have been completely conserved, among which are residues 25-58, including the nucleotide-binding A domain. Especially the C-terminal half (amino acids 195-353), implicated in receptor and effector interactions, is highly conserved (94% sequence identity with murine sequences). This region includes the nucleotide-binding C, G, and I domains, which are identical to cognate motifs of vertebrate G alpha q/11. Like the latter proteins, the Lymnaea G alpha q C-terminus lacks a cysteine that could serve as a substrate for pertussis toxin. In situ hybridization reveals G alpha q-encoding mRNA(s) to be present throughout the CNS. Interestingly, however, close inspection of two identified cell types in the cerebral ganglia, the light-green cells, involved in the regulation of growth and metabolism and the anterior lobe cells which are involved in the control of male aspects of reproduction, indicates that they express the mRNA(s) at significantly different levels. Even within the heterologous cluster of light-green cells there appears to be differential expression of the pertinent mRNA. Such observations have hitherto not been reported for specific cell types occurring in vivo.

摘要

通过分子克隆,我们鉴定出一种软体动物G蛋白α亚基,它属于Gαq家族,在椎实螺(Lymnaea stagnalis)的中枢神经系统(CNS)中表达。推导的蛋白质产物与脊椎动物和无脊椎动物的Gαq / Gα11亚基具有非常高的氨基酸序列同一性(分别为80 - 82%和76 - 77%)。该蛋白质的大部分区域完全保守,其中包括25 - 58位残基,包括核苷酸结合A结构域。特别是参与受体和效应器相互作用的C端一半(氨基酸195 - 353)高度保守(与鼠序列的序列同一性为94%)。该区域包括核苷酸结合C、G和I结构域,它们与脊椎动物Gαq / 11的同源基序相同。与后一种蛋白质一样,椎实螺Gαq的C末端缺少一个可作为百日咳毒素底物的半胱氨酸。原位杂交显示整个中枢神经系统中都存在编码Gαq的mRNA。然而,有趣的是,仔细检查脑神经节中两种已鉴定的细胞类型,即参与生长和代谢调节的浅绿色细胞和参与雄性生殖控制的前叶细胞,发现它们表达的mRNA水平存在显著差异。即使在浅绿色细胞的异源簇中,相关mRNA似乎也存在差异表达。迄今为止,尚未报道过体内特定细胞类型的此类观察结果。

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