Microseeding - a powerful tool for crystallizing proteins complexed with hydrolyzable substrates.

ABSTRACT: Hydrolysis is an often-encountered obstacle in the crystallization of proteins complexed with their substrates. As the duration of the crystallization process, from nucleation to the growth of the crystal to its final size, commonly requires several weeks, non-enzymatic hydrolysis of an "unstable" ligand occurs frequently. In cases where the crystallization conditions exhibit non neutral pH values this hydrolysis phenomenon may be even more pronounced. ChoX, the substrate binding protein of a choline ABC-importer, produced crystals with its substrate acetylcholine after one month. However, these crystals exhibited only choline, an acetylcholine hydrolysis product, in the binding site. To overcome this obstacle we devised a microseeding protocol leading to crystals of ChoX with bound acetyl

SUBMITTER: Oswald C 

PROVIDER: S-EPMC2635726 | biostudies-literature | 2008 Jun

REPOSITORIES: biostudies-literature