Fast identification of folded human protein domains expressed in E. coli suitable for structural analysis.

ABSTRACT:

Background

High-throughput protein structure analysis of individual protein domains requires analysis of large numbers of expression clones to identify suitable constructs for structure determination. For this purpose, methods need to be implemented for fast and reliable screening of the expressed proteins as early as possible in the overall process from cloning to structure determination.

Results

88 different E. coli expression constructs for 17 human protein domains were analysed using high-throughput cloning, purification and folding analysis to obtain candidates suitable for structural analysis. After 96 deep-well microplate expression and automated protein purification, protein domains were directly analysed using 1D 1H-NMR spectroscopy. In addition, analytical hydropho

SUBMITTER: Scheich C 

PROVIDER: S-EPMC516802 | biostudies-literature | 2004 Mar

REPOSITORIES: biostudies-literature