A unified purification method for actin-binding proteins using a TEV-cleavable His-Strep-tag.

ABSTRACT: The actin cytoskeleton governs the dynamic functions of cells, ranging from motility to phagocytosis and cell division. To elucidate the molecular mechanism, in vitro reconstructions of the actin cytoskeleton and its force generation process have played essential roles, highlighting the importance of efficient purification methods for actin-binding proteins. In this study, we introduce a unified purification method for actin-binding proteins, including capping protein (CP), cofilin, ADF, profilin, fascin, and VASP, key regulators in force generation of the actin cytoskeleton. Exploiting a His-Strep-tag combined with a TEV protease cleavage site, we purified these diverse actin-binding proteins through a simple two-column purification process: initial purification through a Strep-Tac

SUBMITTER: Nakajima D 

PROVIDER: S-EPMC11367271 | biostudies-literature | 2024 Dec

REPOSITORIES: biostudies-literature