Characterization of retinal guanylate cyclase-activating protein 3 (GCAP3) from zebrafish to man
Characterization of retinal guanylate cyclase-activating protein 3 (GCAP3) from zebrafish to man
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DOI：
10.1046/j.0953-816x.2001.01835.x
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发表时间：
2002-01-01
影响因子：
3.4
通讯作者：
Palczewski, K
中科院分区：
文献类型：
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作者：
Imanishi, Y;Li, G;Palczewski, K
钙调素样神经元钙结合蛋白（NCBPs）主要在神经元中表达，包含四个功能性和非功能性EF-手钙结合基序的组合。鸟苷酸环化酶激活蛋白1-3（GCAP 1 -3）是最具特征的NCBP亚组，其功能是调节跨膜鸟苷酸环化酶1-2（GC 1 -2）。GCAP和GC在体内的配对取决于细胞表达。因此，我们采用原位杂交和免疫细胞化学方法研究了这些基因在视网膜中的表达。我们的研究结果表明，GCAP 1，GCAP 2，GC 1和GC 2在人类视杆细胞和视锥细胞中表达，而GCAP 3只在视锥细胞中表达。作为广泛修饰的结果，GCAP 3基因在小鼠视网膜中不表达。然而，这种缺乏进化保守性似乎仅限于某些物种，因为我们从硬骨鱼（斑马鱼）视网膜克隆了所有三种GCAP，并将其定位于视杆细胞，短单视锥（GCAP 1 -2）和所有亚型的视锥（GCAP 3）。此外，序列比较和进化的痕迹分析加上不同的GCAP的功能测试，使我们能够确定关键的保守残基，是关键的GCAP结构和功能，并定义类特异性残基的NCBP亚家族。
Calmodulin-like neuronal Ca2+-binding proteins (NCBPs) are expressed primarily in neurons and contain a combination of four functional and nonfunctional EF-hand Ca2+-binding motifs. The guanylate cyclase-activating proteins 1-3 (GCAP1-3), the best characterized subgroup of NCBPs, function in the regulation of transmembrane guanylate cyclases 1-2 (GC1-2). The pairing of GCAPs and GCs in vivo depends on cell expression. Therefore, we investigated the expression of these genes in retina using in situ hybridization and immunocytochemistry. Our results demonstrate that GCAP1, GCAP2, GC1 and GC2 are expressed in human rod and cone photoreceptors, while GCAP3 is expressed exclusively in cones. As a consequence of extensive modification, the GCAP3 gene is not expressed in mouse retina. However, this lack of evolutionary conservation appears to be restricted to only some species as we cloned all three GCAPs from teleost (zebrafish) retina and localized them to rod cells, short single cones (GCAP1-2), and all subtypes of cones (GCAP3). Furthermore, sequence comparisons and evolutionary trace analysis coupled with functional testing of the different GCAPs allowed us to identify the key conserved residues that are critical for GCAP structure and function, and to define class-specific residues for the NCBP subfamilies.