Ando-Akatsuka Y, Saitou M, Hirase T, Kishi M, Sakakibara A, Itoh M, Yonemura S, Furuse M, Tsukita S
Department of Cell Biology, Faculty of Medicine, Kyoto University, Japan.
J Cell Biol. 1996 Apr;133(1):43-7. doi: 10.1083/jcb.133.1.43.
Occludin has been identified from chick liver as a novel integral membrane protein localizing at tight junctions (Furuse, M., T. Hirase, M. Itoh, A. Nagafuchi, S. Yonemura, Sa. Tsukita, and Sh. Tsukita. 1993. J. Cell Biol. 123:1777-1788). To analyze and modulate the functions of tight junctions, it would be advantageous to know the mammalian homologues of occludin and their genes. Here we describe the nucleotide sequences of full length cDNAs encoding occludin of rat-kangaroo (potoroo), human, mouse, and dog. Rat-kangaroo occludin cDNA was prepared from RNA isolated from PtK2 cell culture, using a mAb against chicken occludin, whereas the others were amplified by polymerase chain reaction based on the sequence found around the human neuronal apoptosis inhibitory protein gene. The amino acid sequences of the three mammalian (human, murine, and canine) occludins were very closely related to each other (approximately 90% identity), whereas they diverged considerably from those of chicken and rat-kangaroo (approximately 50% identity). Implications of these data and novel experimental options in cell biological research are discussed.
封闭蛋白是从小鸡肝脏中鉴定出的一种新型整合膜蛋白,定位于紧密连接(Furuse, M., T. Hirase, M. Itoh, A. Nagafuchi, S. Yonemura, Sa. Tsukita, and Sh. Tsukita. 1993. J. Cell Biol. 123:1777 - 1788)。为了分析和调节紧密连接的功能,了解封闭蛋白的哺乳动物同源物及其基因将是有利的。在此,我们描述了编码大鼠袋鼠(长鼻袋鼠)、人类、小鼠和狗的封闭蛋白的全长cDNA的核苷酸序列。大鼠袋鼠封闭蛋白cDNA是从PtK2细胞培养物中分离的RNA制备的,使用针对鸡封闭蛋白的单克隆抗体,而其他的则基于人类神经元凋亡抑制蛋白基因周围发现的序列通过聚合酶链反应进行扩增。三种哺乳动物(人类、小鼠和犬类)封闭蛋白的氨基酸序列彼此非常密切相关(约90%的同一性),而它们与鸡和大鼠袋鼠的氨基酸序列有很大差异(约50%的同一性)。本文讨论了这些数据的意义以及细胞生物学研究中的新实验选择。