Kurobe Tomofumi, Hirono Ikuo, Kondo Hidehiro, Yamashita Michiaki, Aoki Takashi
Laboratory of Genome Science, Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, Konan 4-5-7, Minato, Tokyo 108-8477, Japan.
Fish Shellfish Immunol. 2007 Dec;23(6):1266-74. doi: 10.1016/j.fsi.2007.07.001. Epub 2007 Jul 19.
We isolated and sequenced caspase-10 cDNA and gene from Japanese flounder, Paralichthys olivaceus. The Japanese flounder (JF)-caspase-10 cDNA consisted of 2282 bp and encoded 495 amino acid residues. The characteristic death effector domains (DEDs) of caspases were observed in JF-caspase-10 as well as the three aspartic acid residues (D-186, -382 and -392), which are potential cleavage sites for the large and small subunit structures. The amino acid residue (His-325) and pentapeptide (QACQG), which are involved in catalytic activity, were absolutely conserved in Japanese flounder-caspase-10. JF-caspase-10 gene has a length of 6.6 kb and consists of 11 exons and 10 introns similar to that of human. The strong expression of JF-caspase-10 mRNA was detected in the gills, peripheral blood leukocytes, spleen and posterior kidney, while the weak expression was observed in the head kidney, heart, intestine, skin and stomach. The over-expression analysis of JF-caspase-10 in Japanese flounder cell line HINAE was shown to induce apoptosis 24h post-transfection using TUNEL assay.
我们从日本牙鲆（Paralichthys olivaceus）中分离并测序了caspase-10 cDNA和基因。日本牙鲆（JF）-caspase-10 cDNA由2282个碱基对组成，编码495个氨基酸残基。在JF-caspase-10中观察到了半胱天冬酶特有的死亡效应结构域（DEDs）以及三个天冬氨酸残基（D-186、-382和-392），它们是大小亚基结构的潜在切割位点。参与催化活性的氨基酸残基（His-325）和五肽（QACQG）在日本牙鲆-caspase-10中绝对保守。JF-caspase-10基因长度为6.6 kb，由11个外显子和10个内含子组成，与人类的相似。在鳃、外周血白细胞、脾脏和后肾中检测到JF-caspase-10 mRNA的强表达，而在头肾、心脏、肠道、皮肤和胃中观察到弱表达。使用TUNEL检测法对日本牙鲆细胞系HINAE中的JF-caspase-10进行过表达分析，结果显示转染后24小时可诱导细胞凋亡。
