Phylogeny and expression of carbonic anhydrase-related proteins.
Phylogeny and expression of carbonic anhydrase-related proteins.
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DOI：
10.1186/1471-2199-11-25
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发表时间：
2010-03-31
影响因子：
--
通讯作者：
Parkkila S
中科院分区：
文献类型：
--
作者：
Aspatwar A;Tolvanen ME;Parkkila S
碳酸酐酶（Carbonic anhydrase，CA）是一类存在于多种生物体中的酶，参与多种重要的生物学过程。脊椎动物α-CA家族由16个亚家族组成，其中三个（VIII、X和XI）由非催化蛋白组成。这些被命名为碳酸酐酶相关蛋白（CARP），它们的失活是由于缺乏一个或多个锌结合组氨酸残基。在这项研究中，我们分析和评估的基因编码的CARP在不同的生物体中的分布，并研究其在小鼠组织中的表达，免疫组织化学和实时定量PCR。我们收集了84个序列，其中22个来自我们从基因组数据创建的新的或改进的基因模型。CARP VIII的分布覆盖了脊椎动物和后口动物，而CARP X似乎在动物界中是普遍存在的。CA 10样基因在四足动物和鱼类谱系中有单独的复制历史。我们的系统发育分析表明，重复的CA 10到CA 11只发生在四足动物（发现在哺乳动物，青蛙和蜥蜴），而一个独立的重复的CA 10被发现在鱼类。我们建议将第二种鱼类亚型命名为CA 10 b。免疫组织化学分析显示CARP VIII在小鼠小脑、大脑中有高水平表达，在肺、肝、唾液腺和胃中也有中等水平表达。这些结果还表明在结肠、肾脏和朗格汉斯胰岛中的低表达。CARP X在脑毛细血管和肺中中度表达，在胃和心脏中非常弱。在小脑、大脑、肝脏、胃、小肠、结肠、肾脏和睾丸中观察到CARP XI的阳性信号。此外，实时定量PCR的结果证实了广泛分布的Car 8和Car 11的mRNA，而表达的Car 10的mRNA仅限于额叶皮质，顶叶皮质，小脑，中脑和眼睛。CARP序列在不同物种之间高度保守，所有三种CARP在小鼠脑中均显示高表达，CARP VIII也在几种其他组织中表达。这些发现表明这些蛋白质在哺乳动物中具有重要的功能作用。
Carbonic anhydrases (CAs) are found in many organisms, in which they contribute to several important biological processes. The vertebrate α-CA family consists of 16 subfamilies, three of which (VIII, X and XI) consist of acatalytic proteins. These are named carbonic anhydrase related proteins (CARPs), and their inactivity is due to absence of one or more Zn-binding histidine residues. In this study, we analyzed and evaluated the distribution of genes encoding CARPs in different organisms using bioinformatic methods, and studied their expression in mouse tissues using immunohistochemistry and real-time quantitative PCR. We collected 84 sequences, of which 22 came from novel or improved gene models which we created from genome data. The distribution of CARP VIII covers vertebrates and deuterostomes, and CARP X appears to be universal in the animal kingdom. CA10-like genes have had a separate history of duplications in the tetrapod and fish lineages. Our phylogenetic analysis showed that duplication of CA10 into CA11 has occurred only in tetrapods (found in mammals, frogs, and lizards), whereas an independent duplication of CA10 was found in fishes. We suggest the name CA10b for the second fish isoform. Immunohistochemical analysis showed a high expression level of CARP VIII in the mouse cerebellum, cerebrum, and also moderate expression in the lung, liver, salivary gland, and stomach. These results also demonstrated low expression in the colon, kidney, and Langerhans islets. CARP X was moderately expressed in the cerebral capillaries and the lung and very weakly in the stomach and heart. Positive signals for CARP XI were observed in the cerebellum, cerebrum, liver, stomach, small intestine, colon, kidney, and testis. In addition, the results of real-time quantitative PCR confirmed a wide distribution for the Car8 and Car11 mRNAs, whereas the expression of the Car10 mRNA was restricted to the frontal cortex, parietal cortex, cerebellum, midbrain, and eye. CARP sequences have been strongly conserved between different species, and all three CARPs show high expression in the mouse brain and CARP VIII is also expressed in several other tissues. These findings suggest an important functional role for these proteins in mammals.
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