Cingulin interacts with F‐actin in vitro

FEBS Letters

D'Atri, Fabio¹  Citi, Sandra¹ 

[1] Department of Molecular Biology, University of Geneva, 30 Quai Ernest Ansermet, 1211 Geneva 4, Switzerland

关键词: Cingulin;    Actin;    α-Actinin;    Tight junction;    GST;    glutathione S-transferase;    TJ;    tight junction;   

DOI  :  10.1016/S0014-5793(01)02936-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Cingulin, a M _r 140–160 kDa protein of the cytoplasmic plaque of epithelial tight junctions (TJ), interacts in vitro with TJ proteins and myosin. Here we investigated cingulin interaction with actin, using His-tagged, full-length Xenopus laevis cingulin expressed in insect cells, and glutathione S-transferase (GST) fusion proteins of fragments of cingulin expressed in bacteria. Purified full-length cingulin co-pelleted with F-actin after high speed centrifugation, and promoted the sedimentation of F-actin under low speed centrifugation, suggesting that cingulin is an actin-cross-linking protein. The actin interaction of GST fusion proteins containing fragments of Xenopus cingulin suggested that the F-actin binding site is between residues 101 and 294.

【 授权许可】

Unknown