A technique for high-throughput protein crystallization in ionically cross-linked polysaccharide gel beads for X-ray diffraction experiments.

ABSTRACT: A simple technique for high-throughput protein crystallization in ionically cross-linked polysaccharide gel beads has been developed for contactless handling of crystals in X-ray crystallography. The method is designed to reduce mechanical damage to crystals caused by physical contact between crystal and mount tool and by osmotic shock during various manipulations including cryoprotection, heavy-atom derivatization, ligand soaking, and diffraction experiments. For this study, protein crystallization in alginate and κ-carrageenan gel beads was performed using six test proteins, demonstrating that proteins could be successfully crystallized in gel beads. Two complete diffraction data sets from lysozyme and ID70067 protein crystals in gel beads were collected at 100 K without removing the cry

SUBMITTER: Sugahara M 

PROVIDER: S-EPMC3989300 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature