Premium
Cysteine Mutagenesis to Study the Structure of Claudin‐2 Paracellular Pores
Author(s) -
Angelow Susanne,
Yu Alan S.L.
Publication year - 2009
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1111/j.1749-6632.2009.04038.x
Subject(s) - paracellular transport , claudin , chemistry , cysteine , mutagenesis , biophysics , transcellular , transmembrane protein , microbiology and biotechnology , biochemistry , tight junction , membrane , biology , permeability (electromagnetism) , mutation , gene , receptor , enzyme
The structure and transport mechanism of paracellular pores are only poorly understood. Here we describe for the first time how the substituted cysteine accessibility method (SCAM), previously developed to study transmembrane transport, can be applied to analyze the pathway of paracellular ion permeation. Using stable transfected Madin Darby canine kidney type I cells, induced to express claudin‐2, we show that paracellular cation transport can be blocked by sulfhydryl‐specific methanethiosulfonate (MTS) and that SCAM can be used to identify residues that line paracellular pores.