Meher Akshaya K, Blaber Sachiko I, Lee Jihun, Honjo Ejiro, Kuroki Ryota, Blaber Michael
Department of Biomedical Sciences, Florida State University, Tallahassee, 32306-4300, USA.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009 Nov 1;65(Pt 11):1136-40. doi: 10.1107/S1744309109036987. Epub 2009 Oct 30.
Large-volume protein crystals are a prerequisite for neutron diffraction studies and their production represents a bottleneck in obtaining neutron structures. Many protein crystals that permit the collection of high-resolution X-ray diffraction data are inappropriate for neutron diffraction owing to a plate-type morphology that limits the crystal volume. Human fibroblast growth factor 1 crystallizes in a plate morphology that yields atomic resolution X-ray diffraction data but has insufficient volume for neutron diffraction. The thin physical dimension has been identified as corresponding to the b cell edge and the X-ray structure identified a solvent-mediated crystal contact adjacent to position Glu81 that was hypothesized to limit efficient crystal growth in this dimension. In this report, a series of mutations at this crystal contact designed to both reduce side-chain entropy and replace the solvent-mediated interface with direct side-chain contacts are reported. The results suggest that improved crystal growth is achieved upon the introduction of direct crystal contacts, while little improvement is observed with side-chain entropy-reducing mutations alone.
大体积蛋白质晶体是中子衍射研究的前提条件,其生产是获得中子结构的一个瓶颈。许多能够收集高分辨率X射线衍射数据的蛋白质晶体由于其平板状形态限制了晶体体积,因而不适用于中子衍射。人成纤维细胞生长因子1以平板形态结晶,可产生原子分辨率的X射线衍射数据,但体积不足以用于中子衍射。已确定其薄的物理尺寸对应于b晶胞边缘,并且X射线结构确定了在Glu81位置附近存在溶剂介导的晶体接触,据推测这限制了该维度上晶体的有效生长。在本报告中,报道了针对该晶体接触处的一系列突变,这些突变旨在既降低侧链熵,又用直接的侧链接触取代溶剂介导的界面。结果表明,引入直接的晶体接触可实现晶体生长的改善,而单独的降低侧链熵的突变几乎没有观察到改善。