Identifying protein construct variants with increased crystallization propensity--a case study.

ABSTRACT: This study describes an efficient multiparallel automated workflow of cloning, expression, purification, and crystallization of a large set of construct variants for isolated protein domains aimed at structure determination by X-ray crystallography. This methodology is applied to MAPKAP kinase 2, a key enzyme in the inflammation pathway and thus an attractive drug target. The study reveals a distinct subset of truncation variants with improved crystallization properties. These constructs distinguish themselves by increased solubility and stability during a parallel automated multistep purification process including removal of the recombinant tag. High-throughput protein melting point analysis characterizes this subset of constructs as particularly thermostable. Both parallel purification s

SUBMITTER: Malawski GA 

PROVIDER: S-EPMC2242438 | biostudies-literature | 2006 Dec

REPOSITORIES: biostudies-literature