Identification of gelsolin as an actin regulatory component in a Triton insoluble law density fraction (raft) of newborn bovine brain

A membrane microdomain enriched in cholesterol and glycosphingolipids, or so called 'raft' region, was found to contain many signal transducing proteins such as GPI-anchored cell adhesion molecules, trimeric G proteins, and protein tyrosine kinases. In previous studies, we showed that the raft region obtained from rat brain contains two cytoskeletal proteins, tubulin and actin, as the major components in addition to these signal transducing proteins. In this study, to know the biochemical mechanisms regulating the cytoskeletal organization in this region, actin regulatory activities in raft were surveyed. We found the presence of a Ca2+-dependent actin nucleation promoting activity in raft. The solubilization and column fractionation of this activity combined with western blotting and immunoprecipitation showed that gelsolin is one of the actin regulatory proteins in raft. (C) 2000 Published by Elsevier Science Ireland Ltd and the Japan Neuroscience Society. All rights reserved.