Growth of Large Protein Crystals by Top-Seeded Solution Growth Together with the Floating and Solution-Stirring Technique

摘要

Neutron protein crystallography is expected to yield a more precise understanding of the protein structure by visualizing hydrogen atoms and hydration water molecules. However, relatively few protein structures have been analyzed by neutron diffraction, in part because very large crystals of the target proteins are required in order to compensate for the weak flux of the available neutron beam. Here, we report the crystal growth of hen egg white lysozyme (HEWL) by the top-seeded solution growth with the floating and solution-stirring method (TSSG-FAST), the first utilized for protein crystallization. A series of crystal-growth experiments demonstrated that TSSG-FAST is an efficient strategy for rapidly obtaining large single crystals and effectively preventing polycrystallization of the seed crystal. Furthermore, the shape of the HEWL crystals was found to be strongly influenced by the orientation of the seed crystal, Suggesting that the shape of the HEWL crystal may be controlled by TSSG-FAST. It is anticipated that this new approach will expand the limits on the growth of large protein crystals for neutron protein crystallography.