Inagaki N, Mizuta M, Seino S
Division of Molecular Medicine, Chiba University School of Medicine.
J Biochem. 1994 Aug;116(2):239-42. doi: 10.1093/oxfordjournals.jbchem.a124512.
Rab3A, a ras p21-related small GTP-binding protein, is implicated in the exocytosis of neurotransmitters. Recently, Rabphilin-3A, a putative target protein for Rab3A, was identified and its cDNA has been cloned from bovine brain. In this study, we isolated a cDNA encoding a mouse Rabphilin-3A homolog from the insulin-secreting cell line, MIN6. Mouse Rabphilin-3A is a protein of 681 amino acids exhibiting overall 88.5% identity with bovine Rabphilin-3A. The amino acid identity between mouse and bovine Rabphilin-3A is highest in their carboxyl-terminal halves (97.8% identity) and amino-termini (93.0% identity), which contain the region of the two internal repeats homologous to the regulatory domain (C2 domain) of protein kinase C and the putative Rab3A-binding region, respectively. RNA blot analysis revealed that Rabphilin-3A mRNA is expressed in endocrine and hormone-secreting clonal cells, including rat adrenal glands, MIN6, the hamster insulin-secreting cell line, HIT-T15, and the rat catecholamine-secreting cell line, PC12, as well as rat brain. These results suggest that Rabphilin-3A might be involved in the exocytosis of secretory vesicles in hormone-secreting cells as well as in neurons.
Rab3A是一种与ras p21相关的小GTP结合蛋白,与神经递质的胞吐作用有关。最近,Rabphilin-3A作为Rab3A的一种假定靶蛋白被鉴定出来,其cDNA已从牛脑中克隆出来。在本研究中,我们从胰岛素分泌细胞系MIN6中分离出编码小鼠Rabphilin-3A同源物的cDNA。小鼠Rabphilin-3A是一种由681个氨基酸组成的蛋白质,与牛Rabphilin-3A的整体同源性为88.5%。小鼠和牛Rabphilin-3A之间的氨基酸同源性在它们的羧基末端(同源性为97.8%)和氨基末端(同源性为93.0%)最高,这两个区域分别包含与蛋白激酶C的调节结构域(C2结构域)同源的两个内部重复区域和假定的Rab3A结合区域。RNA印迹分析显示,Rabphilin-3A mRNA在包括大鼠肾上腺、MIN6、仓鼠胰岛素分泌细胞系HIT-T15、大鼠儿茶酚胺分泌细胞系PC12以及大鼠脑在内的内分泌和激素分泌克隆细胞中表达。这些结果表明,Rabphilin-3A可能参与激素分泌细胞以及神经元中分泌囊泡的胞吐作用。