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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Cloning and expression of a second photoreceptor-specific membrane retina guanylyl cyclase (RetGC), RetGC-2.
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Cloning and expression of a second photoreceptor-specific membrane retina guanylyl cyclase (RetGC), RetGC-2.

机译：第二个光感受器特异性膜视网膜鸟苷酸环化酶（RetGC）RetGC-2的克隆和表达。

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摘要

One of the membrane guanylyl cyclases (GCs), RetGC, is expressed predominantly in photoreceptors. No extracellular ligand has been described for RetGC, but it is sensitive to activation by a soluble 24-kDa protein (p24) and is inhibited by Ca2+. This enzyme is, therefore, thought to play a role in resynthesizing cGMP for photoreceptor recovery or adaptation. By screening a human retinal cDNA library at low stringency with the cytoplasmic domains from four cyclases, we cloned cDNAs encoding a membrane CG that is most closely related to RetGC. We have named this GC RetGC-2, and now term the initially described RetGC RetGC-1. By in situ hybridization, mRNA encoding RetGC-2 is found only in the outer nuclear layer and inner segments of photoreceptor cells. By using synthetic peptide antiserum specific for each RetGC subtype, RetGC-2 can be distinguished from RetGC-1 as a slightly smaller protein in immunoblots of bovine rod outer segments. Membrane GC activity of recombinant RetGC-2 expressed in human embryonic kidney 293 cells is stimulated by the activator p24 and is inhibited by Ca2+ with an EC50 value of 50-100 nM. Our data reveal a previously unappreciated diversity of photoreceptor GCs.
机译：膜鸟苷基环化酶（GC）之一RetGC主要在感光器中表达。没有针对RetGC的细胞外配体的描述，但它对可溶性24 kDa蛋白（p24）的激活敏感，并被Ca2 +抑制。因此，认为该酶在用于光受体恢复或适应的cGMP的再合成中起作用。通过筛选来自四个环化酶的胞质结构域的低严格性人类视网膜cDNA文库，我们克隆了编码与RetGC最紧密相关的膜CG的cDNA。我们已将此GC命名为RetGC-2，现在将其称为最初描述的RetGC RetGC-1。通过原位杂交，仅在感光细胞的外核层和内部节段中发现了编码RetGC-2的mRNA。通过使用对每种RetGC亚型具有特异性的合成肽抗血清，RetGC-2可与RetGC-1区别开来，它是牛杆外部区段免疫印迹中稍小的蛋白质。在人类胚胎肾293细胞中表达的重组RetGC-2的膜GC活性被激活剂p24刺激，并被Ca2 +抑制，EC50值为50-100 nM。我们的数据揭示了感光体GC以前未曾意识到的多样性。

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