CoESPRIT: a library-based construct screening method for identification and expression of soluble protein complexes.

ABSTRACT: Structural and biophysical studies of protein complexes require multi-milligram quantities of soluble material. Subunits are often unstable when expressed separately so co-expression strategies are commonly employed since in vivo complex formation can provide stabilising effects. Defining constructs for subunit co-expression experiments is difficult if the proteins are poorly understood. Even more problematic is when subunit polypeptide chains co-fold since individually they do not have predictable domains. We have developed CoESPRIT, a modified version of the ESPRIT random library construct screen used previously on single proteins, to express soluble protein complexes. A random library of target constructs is screened against a fixed bait protein to identify stable complexes. In a proof-

SUBMITTER: An Y 

PROVIDER: S-EPMC3043051 | biostudies-literature | 2011 Feb

REPOSITORIES: biostudies-literature