Purification of modified mammalian actin isoforms for in vitro reconstitution assays.

ABSTRACT: In vitro reconstitution assays using purified actin have greatly improved our understanding of cytoskeletal dynamics and their regulation by actin-binding proteins. However, early purification methods consisted of harsh conditions to obtain pure actin and often did not include correct maturation and obligate modification of the isolated actin monomers. Novel insights into the folding requirements and N-terminal processing of actin as well as a better understanding of the interaction of actin with monomer sequestering proteins such as DNaseI, profilin and gelsolin, led to the development of more gentle approaches to obtain pure recombinant actin isoforms with known obligate modifications. This review summarizes the approaches that can be employed to isolate natively folded endogenous and re

SUBMITTER: Kast DJ 

PROVIDER: S-EPMC10872616 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature