A gelsolin-like Ca2+-dependent actin-binding domain in villin.

Villin is an actin-binding protein of relative molecular mass (Mr) 95,000 found in the core bundle of microfilaments in brush border microvilli from intestine. In physiological calcium concentrations (less than 1 microM), villin crosslinks actin filaments into bundles. However, in free calcium concentrations (greater than 1 microM), villin severs actin filaments into short pieces. To understand how villin can sever and bundle actin filaments, we are studying the molecular basis of villin-actin binding interactions by identifying important actin-binding domains in villin. Here, we report the purification and preliminary characterization of a 44,000-Mr fragment of villin which contains a calcium-dependent actin-severing activity. In addition, the partial amino-acid sequence from the amino terminus of this fragment reveals homology with a 16-residue region near the amino terminus of gelsolin, an actin-severing protein found in many cells and sera. The sequence homology suggests a common structural basis for the calcium-regulated actin-severing properties shared by villin and gelsolin.