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首页> 外文期刊>Biochemistry >Identification of a guanylyl cyclase-activating protein-binding site within the catalytic domain of retinal guanylyl cyclase 1.
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Identification of a guanylyl cyclase-activating protein-binding site within the catalytic domain of retinal guanylyl cyclase 1.

机译：鉴定视网膜鸟苷酸环化酶1催化域内的鸟苷酸环化酶激活蛋白结合位点。

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摘要

Regulation of cAMP and cGMP production is a fundamental step in a broad range of signal transduction systems, including phototransduction. To identify regions within photoreceptor guanylyl cyclase 1 (GC1) that interact with GC-activating proteins (GCAPs), we synthesized the intracellular fragment of GC1, residues 491-1110, as a set of 15 amino acid long, partially overlapping peptides on the surface of individual pins arranged in a microtiter plate format. This pin assay identified 8 peptides derived from different regions of the GC1 intracellular domain that bind GCAPs. Peptide variants containing these sequences were synthesized as free peptides and tested for their ability to inhibit GC1 stimulation by GCAPs. A free peptide,968GTFRMRHMPEVPVRIRIG, from the catalytic domain of GC1 was the strongest inhibitor of GCAP1/GCAP2-mediated activation. In native GC1, this polypeptide fragment is likely to form a loop between alpha-helix 3 and beta-strand 4. When this region in GC1 was replaced by the corresponding sequence of GCAP-insensitive GC type A, GCAPs did not stimulate the GC1 mutant. The corresponding loops in related adenylyl cyclase (AC) are involved in the activating and inhibiting interactions with Gs alpha and Gi alpha, respectively. Thus, despite interacting with different activating proteins, both AC and GC activity may be modulated through their respective regions within catalytic domains.
机译：调节cAMP和cGMP的产生是包括光转导在内的各种信号转导系统中的基本步骤。为了鉴定感光受体鸟苷酰环化酶1（GC1）中与GC激活蛋白（GCAP）相互作用的区域，我们合成了GC1的细胞内片段，残基491-1110，为一组15个氨基酸长，部分重叠的表面肽以微量滴定板形式排列的单个销钉。该pin测定法鉴定了8种来自GC1胞内域不同区域的结合GCAP的肽。将包含这些序列的肽变体合成为游离肽，并测试其抑制GCAP刺激GC1的能力。来自GC1催化域的游离肽968GTFRMRHMPEVPVRIRIG是GCAP1 / GCAP2介导的激活的最强抑制剂。在天然GC1中，此多肽片段很可能在α-螺旋3和β-链4之间形成环。当GC1中的该区域被GCAP不敏感的A型GC对应序列取代时，GCAP不会刺激GC1突变体。相关的腺苷酸环化酶（AC）中的相应环分别参与与Gs alpha和Gi alpha的激活和抑制相互作用。因此，尽管与不同的活化蛋白相互作用，但是AC和GC活性都可以通过它们在催化域内的相应区域来调节。

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