Molecular cloning of a murine glycerol-3-phosphate acyltransferase-like protein 1 (xGPAT1)
Molecular cloning of a murine glycerol-3-phosphate acyltransferase-like protein 1 (xGPAT1)
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DOI：
10.1007/s11010-006-9321-5
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发表时间：
2007-03-01
影响因子：
4.3
通讯作者：
Nakaya, Yutaka
中科院分区：
文献类型：
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作者：
Harada, Nagakatsu;Hara, Sayuri;Nakaya, Yutaka
克隆了一种新的小鼠甘油-3-磷酸酰基转移酶样蛋白1（命名为xGPAT1）。小鼠xGPAT1基因位于小鼠2号染色体上，长度为bbbb19 kb，由至少23个外显子组成。该蛋白在氨基酸水平上与小鼠线粒体GPAT （mtGPAT）相同32%，相似72%。测序分析证实，xGPAT1具有2403 bp的开放阅读框（ORF），编码一个801个氨基酸的蛋白，估计分子质量为89.1 kDa。推导出的xGPAT1蛋白的亲水图与mtGPAT蛋白高度相似。利用cDNA末端的5'快速扩增，获得两个交替的未翻译外显子1 （1a和b）异构体，产生变异xGPAT1-v1和xGPAT1-v2。xGPAT1-v1表达于小鼠心、肝、脾、肾和小鼠髓内集管3 （mIMCD3）细胞，xGPAT1-v2表达于小鼠肝、脾、肾、白色和棕色脂肪组织以及3T3-L1脂肪前和脂肪后细胞。xGPAT1分布在膜组分中，表位标记的xGPAT1在中国仓鼠卵巢(CHO)-K1细胞中表达时显示出GPAT活性。
A novel murine glycerol-3-phosphate acyltransferase-like protein 1 (named xGPAT1) has been cloned. The mouse xGPAT1 gene is located on mouse Chromosome 2, spans > 19 kb, and consists of at least 23 exons. The protein is 32% identical and 72% similar to mouse mitochondrial GPAT (mtGPAT) on the amino acid level. Sequencing analysis confirmed that xGPAT1 has a 2403-bp open reading frame (ORF) that encodes an 801-amino acid protein with an estimated molecular mass of 89.1 kDa. A hydropathy plot of the deduced xGPAT1 protein showed a high degree of similarity with that of the mtGPAT protein. Using 5'-rapid amplification of cDNA ends, two alternate, untranslated exon 1 (1a and b) isoforms were obtained, generating variants xGPAT1-v1 and xGPAT1-v2. xGPAT1-v1 is expressed in mouse heart, liver, spleen, kidney and murine inner medullary collecting duct 3 (mIMCD3) cells, while xGPAT1-v2 is expressed in mouse liver, spleen, kidney, white and brown adipose tissues and 3T3-L1 pre- and post-adipocytes. xGPAT1 was distributed in the membrane fraction and showed GPAT activity when epitope-tagged xGPAT1 was expressed in Chinese hamster ovary (CHO)-K1 cells.