Molecular analysis of the tight junction

Abstract

The tight junction (zonula occludens) constitutes a selectively permeable barrier in the paracellular pathway of most epithelia. It is also thought to play a role in the maintenance of the cell surface compositional asymmetry characteristic of epithelial cells. The identification of ZO-1 and cingulin, the first two proteins found to be exclusively associated with the tight junction, permits novel investigations of this important epithelial cell structure at the biochemical level.ZO-1 is a high molecular weight polypeptide (>200 kD) found at the tight junctions of a variety of epithelia as well as endothelia, and ultrastructural localization studies on isolated liver plasma membranes indicate that this molecule is clustered at the points of membrane contact on the cytoplasmic surface of the junction. Physical analysis demonstrates ZO-1 to be an elongated, monomeric, phosphorylated protein, peripherally associated with the junctional membrane. Cingulin was originally isolated from chicken intestine, and, like ZO-1, is a peripheral membrane component of the junction which exhibits an elongated shape. Antibodies directed against this molecule show two primary bands at 140 kD and 108 kD on immunoblots of several epithelial tissues, although the relationship between these two elements remains undefined. Little information currently exists regarding the relationship of ZO-1 and cingulin to each other or to tight junction structure or function. We report here a comparison of the immunocytochemical properties of these junctional components as well as an examination of the phosphorylation state of ZO-1.