Successful protein cryocooling for powder diffraction

Abstract

It is shown that samples of microcrystalline tetragonal chicken egg white lysozyme can be effectively cryoprotected for high-resolution synchrotron X-ray powder diffraction studies at 100 K. The survival of the powder in the beam is increased by a factor of around 30. Thus, a high-quality powder diffraction pattern could be collected at 100 K, which attains a resolution ofdmin≃ 2.6 Å, significantly better than the previous limit of ∼3.27 Å at room temperature, despite a smaller volume of sample. Systematic variations of the concentration and type of cryoprotectant agent show that the lattice microstrains that accompany cooling, and degrade the quality of the powder diffraction data by broadening the diffraction peaks, are caused by a collapse in the volume of the crystalline unit cell.