Multi-PDZ domain protein 1 (MUPP1) is concentrated at tight junctions through its possible interaction with claudin-1 and junctional adhesion molecule.
Claudins, most of which end in valine at their COOH termini, constitute tight junction (TJ) strands, suggesting that TJ strands strongly attract PDZ-containing proteins. Indeed, ZO-1, -2, and -3, each of which contains three PDZ domains, were shown to directly bind to claudins. Using the yeast two-hybrid system, we identified ZO-1 and MUPP1 (multi-PDZ domain protein 1) as binding partners for the COOH terminus of claudin-1. MUPP1 has been identified as a protein that contains 13 PDZ domains, but it has not been well characterized. In vitro binding assays with recombinant MUPP1 confirmed the interaction between MUPP1 and claudin-1 and identified PDZ10 as the responsible domain for this interaction. A polyclonal antibody specific for MUPP1 was then generated. Immunofluorescence confocal microscopy as well as immunoelectron microscopy with this antibody revealed that in polarized epithelial cells MUPP1 was exclusively concentrated at TJs. Furthermore, in vitro binding and transfection experiments showed that junctional adhesion molecule, another TJ adhesion molecule, also bound to the PDZ9 domain of MUPP1. These findings suggested that MUPP1 is concentrated at TJs in epithelial cells through its binding to claudin and junctional adhesion molecule and that it may function as a multivalent scaffold protein that recruits various proteins to TJs.
Hamazaki
Yoko
Y
Department of Cell Biology, Faculty of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan.
Itoh
Masahiko
M
Sasaki
Hiroyuki
H
Furuse
Mikio
M
Tsukita
Shoichiro
S
D000818:Animals; D002352:Carrier Proteins / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D016764:Cell Polarity; D062445:Claudin-1; D004847:Epithelial Cells / Q000737:chemistry; D062726:Junctional Adhesion Molecules; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D019108:Tight Junctions / Q000378:metabolism*
D002352:Carrier Proteins; D015815:Cell Adhesion Molecules; D062445:Claudin-1; C568113:Cldn1 protein, mouse; D062726:Junctional Adhesion Molecules; D008565:Membrane Proteins; C491750:Mpdz protein, mouse