Warkentin Matthew, Berejnov Viatcheslav, Husseini Naji S, Thorne Robert E
Physics Department, Cornell University, Ithaca, NY 14853, USA.
J Appl Crystallogr. 2006 Dec 1;39(6):805-811. doi: 10.1107/S0021889806037484.
When samples having volumes characteristic of protein crystals are plunge cooled in liquid nitrogen or propane, most cooling occurs in the cold gas layer above the liquid. By removing this cold gas layer, cooling rates for small samples and modest plunge velocities are increased to 1.5 × 10(4) K s(-1), with increases of a factor of 100 over current best practice possible with 10 μm samples. Glycerol concentrations required to eliminate water crystallization in protein-free aqueous mixtures drop from ∼28% w/v to as low as 6% w/v. These results will allow many crystals to go from crystallization tray to liquid cryogen to X-ray beam without cryoprotectants. By reducing or eliminating the need for cryoprotectants in growth solutions, they may also simplify the search for crystallization conditions and for optimal screens. The results presented here resolve many puzzles, such as why plunge cooling in liquid nitrogen or propane has, until now, not yielded significantly better diffraction quality than gas-stream cooling.
当具有蛋白质晶体特征体积的样品在液氮或丙烷中骤冷时,大部分冷却发生在液体上方的冷气体层中。通过去除该冷气体层,小样品在适度骤冷速度下的冷却速率可提高到1.5×10⁴ K s⁻¹,与目前使用10μm样品的最佳做法相比提高了100倍。在无蛋白质的水性混合物中消除水结晶所需的甘油浓度从约28% w/v降至低至6% w/v。这些结果将使许多晶体无需冷冻保护剂就能从结晶盘转移到液态冷冻剂再到X射线束。通过减少或消除生长溶液中对冷冻保护剂的需求,它们还可能简化结晶条件和最佳筛选的搜索。此处呈现的结果解决了许多谜团,例如为什么迄今为止在液氮或丙烷中骤冷并未产生比气流冷却明显更好的衍射质量。