Direct association of occludin with ZO-1 and its possible involvement in the localization of occludin at tight junctions.

• DOI: 10.1083/jcb.127.6.1617
• Publication Date: 2004-05-15T00:22:22Z
• Authors: M Furuse; M Itoh; T Hirase; A Nagafuchi; S Yonemura; S Tsukita; S Tsukita
• Keywords: Occludin

Occludin is an integral membrane protein localizing at tight junctions (TJ) with four transmembrane domains and a long COOH-terminal cytoplasmic domain (domain E) consisting of 255 amino acids. Immunofluorescence laser scan microscopy revealed that chick full-length occludin introduced into human bovine epithelial cells was correctly delivered to incorporated preexisting TJ. Further transfection studies various deletion mutants showed the E, especially its approximately 150 acids E358/504), necessary for localization Secondly, E expressed in Escherichia coli as fusion glutathione-S-transferase, this shown be specifically bound complex ZO-1 (220 kD) ZO-2 (160 among peripheral proteins. In vitro binding analyses using glutathione-S-transferase proteins narrowed down sequence ZO-1/ZO-2 association E358/504. Furthermore, region directly associated recombinant produced E. coli. We concluded itself can localize TJ associate ZO-1. The coincidence suggests underlying cytoskeletons through required localized