Bon C, Dianoux A J, Ferrand M, Lehmann M S
Institut Laue Langevin, B.P.156, 38042 Grenoble Cedex 9, France.
Biophys J. 2002 Sep;83(3):1578-88. doi: 10.1016/S0006-3495(02)73927-1.
This paper reports an incoherent quasielastic neutron scattering study of the single particle, diffusive motions of water molecules surrounding a globular protein, the hen egg-white lysozyme. For the first time such an analysis has been done on protein crystals. It can thus be directly related and compared with a recent structural study of the same sample. The measurement temperature ranged from 100 to 300 K, but focus was on the room temperature analysis. The very good agreement between the structural and dynamical studies suggested a model for the dynamics of water in triclinic crystals of lysozyme in the time range approximately 330 ps and at 300 K. Herein, the dynamics of all water molecules is affected by the presence of the protein, and the water molecules can be divided into two populations. The first mainly corresponds to the first hydration shell, in which water molecules reorient themselves fivefold to 10-fold slower than in bulk solvent, and diffuse by jumps from hydration site to hydration site. The long-range diffusion coefficient is five to sixfold less than for bulk solvent. The second group corresponds to water molecules further away from the surface of the protein, in a second incomplete hydration layer, confined between hydrated macromolecules. Within the time scale probed they undergo a translational diffusion with a self-diffusion coefficient reduced approximately 50-fold compared with bulk solvent. As protein crystals have a highly crowded arrangement close to the packing of macromolecules in cells, our conclusion can be discussed with respect to solvent behavior in intracellular media: as the mobility is highest next to the surface, it suggests that under some crowding conditions, a two-dimensional motion for the transport of metabolites can be dominant.
本文报道了一项关于围绕球状蛋白——鸡蛋清溶菌酶的水分子单粒子扩散运动的非相干准弹性中子散射研究。首次对蛋白质晶体进行了此类分析。因此,它可以直接与对同一样品的近期结构研究相关联并进行比较。测量温度范围为100至300K,但重点是室温分析。结构和动力学研究之间的良好一致性表明了在大约330皮秒的时间范围内以及300K温度下,溶菌酶三斜晶体中水的动力学模型。在此,所有水分子的动力学都受到蛋白质存在的影响,并且水分子可分为两类。第一类主要对应于第一水化层,其中水分子重新定向的速度比在本体溶剂中慢5至10倍,并通过从一个水化位点跳跃到另一个水化位点进行扩散。其长程扩散系数比本体溶剂小5至6倍。第二类对应于远离蛋白质表面的水分子,处于第二个不完整水化层中,被限制在水化的大分子之间。在所探测的时间尺度内,它们进行平移扩散,其自扩散系数与本体溶剂相比降低了约50倍。由于蛋白质晶体具有与细胞中大分子堆积相近的高度密集排列,我们的结论可以就细胞内介质中的溶剂行为进行讨论:由于表面附近的流动性最高,这表明在某些拥挤条件下,代谢物传输的二维运动可能占主导。