Actin co-sedimentation assay; for the analysis of protein binding to F-actin.
The actin cytoskeleton within the cell is a network of actin filaments that allows the movement of cells and cellular processes, and that generates tension and helps maintains cellular shape. Although the actin cytoskeleton is a rigid structure, it is a dynamic structure that is constantly remodeling. A number of proteins can bind to the actin cytoskeleton. The binding of a particular protein to F-actin is often desired to support cell biological observations or to further understand dynamic processes due to remodeling of the actin cytoskeleton. The actin co-sedimentation assay is an in vitro assay routinely used to analyze the binding of specific proteins or protein domains with F-actin. The basic principles of the assay involve an incubation of the protein of interest (full length or domain of) with F-actin, ultracentrifugation step to pellet F-actin and analysis of the protein co-sedimenting with F-actin. Actin co-sedimentation assays can be designed accordingly to measure actin binding affinities and in competition assays.
Srivastava
Jyoti
J
Department of Cell and Tissue Biology, University of California, San Francisco, USA.
Barber
Diane
D
D000199:Actins / Q000032:analysis / Q000378:metabolism*; D003599:Cytoskeleton / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D011485:Protein Binding; D011506:Proteins / Q000032:analysis* / Q000378:metabolism; D014461:Ultracentrifugation / Q000379:methods*
D000199:Actins; D011506:Proteins