Premium
Focal adhesions regulate endothelial junctions via myosin light chain kinase during neutrophil‐induced hyper permeability
Author(s) -
Reynolds Jason J.,
Beard Richard S.,
Haines Ricci J.,
Malinin Nikolay L.,
Wu Mack H.,
Yuan Sarah Y.
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.896.12
Subject(s) - adherens junction , myosin light chain kinase , focal adhesion , microbiology and biotechnology , vascular permeability , endothelial stem cell , inflammation , tight junction , myosin , phosphorylation , chemistry , biology , cadherin , immunology , biochemistry , cell , in vitro , endocrinology
Inflammatory microvascular permeability is mediated largely by neutrophil activation, endothelial transmigration, and release of permeability inducing factors. The dysfunction resulting from the following fluid and protein leak can exacerbate inflammation under pathological conditions. Given the importance of the endothelial barrier in these processes, we investigated an alternative role of non‐muscle myosin light chain kinase (nmMLCK) in neutrophil‐stimulated endothelial permeability. We tested the hypothesis that nmMLCK has substrates other than myosin light chain, and that these targets play a role in the regulation of permeability by neutrophil stimulation. Using Phos‐Tag electrophoresis, we report that nmMLCK is involved in the phosphorylation of proteins from adherens junctions (β‐catenin) as well as focal adhesions (Focal adhesion kinase [FAK], Kindlin 2), and that inhibition of either nmMLCK or FAK can rescue the junction disassembly observed with neutrophil stimulation. This indicates a novel role of FAK in the disassembly of adherens junctions during neutrophil stimulation of endothelial cells. These findings help establish the intracellular molecular pathways underlying neutrophil stimulated endothelial permeability and may help identify therapeutic targets for inflammation‐associated vascular leak and edema. Supported by NIH R01 HL061507, GM097270, HL070752, HL096640.