Reconstitution and protein composition analysis of endocytic actin patches.

ABSTRACT:

Background

Clathrin-actin-mediated endocytosis in yeast involves the progressive assembly of at least 60 different proteins at cortical sites. More than half of these proteins are involved in the assembly of a branched network of actin filaments to provide the forces required for plasma membrane invagination.

Results

To gain insights into the regulation of endocytic actin patch dynamics, we developed an in vitro actin assembly assay using microbeads functionalized with the nucleation promoting factor (NPF) Las17 (yeast WASP). When incubated in a yeast extract, these beads assembled actin networks, and a significant fraction became motile. Multidimensional protein identification technology (MudPIT) showed that the recruitment of actin-binding proteins to these Las17-derived a

SUBMITTER: Michelot A 

PROVIDER: S-EPMC2998891 | biostudies-literature | 2010 Nov

REPOSITORIES: biostudies-literature