Comparative molecular analysis of evolutionarily distant glyceraldehyde-3-phosphate dehydrogenase from Sardina pilchardus and Octopus vulgaris
Comparative molecular analysis of evolutionarily distant glyceraldehyde-3-phosphate dehydrogenase from Sardina pilchardus and Octopus vulgaris
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DOI：
10.1093/abbs/gmq103
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发表时间：
2010-12-01
影响因子：
3.7
通讯作者：
Soukri, Abdelaziz
中科院分区：
文献类型：
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作者：
Baibai, Tarik;Oukhattar, Laila;Soukri, Abdelaziz
依赖于NAD(+)的胞浆甘油醛-3-磷酸脱氢酶(GAPDH，EC 1.2.1.12)是从两个海洋物种的肌肉中提纯的，它们分别是沙丁鱼的骨骼肌和章鱼不可压缩的手臂肌肉。比较生化研究表明，它们的亚基分子质量和等电点值不同。利用从高度保守的蛋白质基序设计的简并引物，通过聚合酶链式反应获得了与沙丁鱼和章鱼GAPC基因内部区域相对应的部分c DNA序列。推导的氨基酸序列的比对被用来建立两个酶的活性部位的3D结构以及沙丁鱼和章鱼酶的系统发育关系。到目前为止，这两种酶分别是硬骨鱼和头足类动物中最早的两种GAPDH。有趣的是，系统发育分析表明，沙丁鱼GAPDH与其他脊椎动物的典型酶属于一个簇，而章鱼GAPDH与其他软体动物的序列在一个更接近细菌和真菌同源物的遥远的基本组装中聚集在一起，从而暗示了它们不同的进化情景。
The NAD(+)-dependent cytosolic glyceraldehyde-3-phosphate dehydrogenase (GAPDH, EC 1.2.1.12), which is recognized as a key to central carbon metabolism in glycolysis and gluconeogenesis and as an important allozymic polymorphic biomarker, was purified from muscles of two marine species: the skeletal muscle of Sardina pilchardus Walbaum (Teleost, Clupeida) and the incompressible arm muscle of Octopus vulgaris (Mollusca, Cephalopoda). Comparative biochemical studies have revealed that they differ in their subunit molecular masses and in pI values. Partial cDNA sequences corresponding to an internal region of the GapC genes from Sardina and Octopus were obtained by polymerase chain reaction using degenerate primers designed from highly conserved protein motifs. Alignments of the deduced amino acid sequences were used to establish the 3D structures of the active site of two enzymes as well as the phylogenetic relationships of the sardine and octopus enzymes. These two enzymes are the first two GAPDHs characterized so far from teleost fish and cephalopod, respectively. Interestingly, phylogenetic analyses indicated that the sardina GAPDH is in a cluster with the archetypical enzymes from other vertebrates, while the octopus GAPDH comes together with other molluscan sequences in a distant basal assembly closer to bacterial and fungal orthologs, thus suggesting their different evolutionary scenarios.