晶胞接种法在肌动蛋白和木聚糖酶结晶中的应用

Application of the Seeding Technique in the Crystallization of Actin and Xylanase

晶胞接种法(seeding technique)包括微晶胞接种法(micro-seeding technique)和大晶胞接种法(macro-seeding technique).微晶胞接种法主要用于改善晶体的衍射质量.大晶胞接种法主要用于得到大体积晶体以提高衍射分辨率,尤其是晶体的中子衍射分辨率.本文采用连续多次微晶胞接种法极大地改进了肌动蛋白(actin)晶体的衍射质量,获得的单晶X-光衍射分辨率达到1.9.本文丰富了大晶胞接种法的内容,以此技术获得了体积达到2立方毫米的木聚糖酶(xylanase)巨大单晶,其中子衍射分辨率达到了2.0,从而为它的第一个中子衍射晶体结构的解析奠定了坚实的基础.

The seeding techniques include the micro- and the macro-seeding technique, where the former is mostly applied to improve diffraction quality, and the latter is used to obtain larger crystals for enhanced diffraction resolution that is important in neutron protein crystallography. We applied micro- seeding technique to obtain an actin crystal with 1.9 A resolution by X-ray. The macro-seeding technique was attempted and obtained a giant xylanase crystal of 2 mm3 with 2.0 A resolution under the neutron beam. It may be further used for the resolution of the first neutron crystal structure of xvlanase.