Development of protein seed crystals reinforced by high-strength hydrogels
Development of protein seed crystals reinforced by high-strength hydrogels
复制标题
高强度水凝胶增强蛋白质晶种的开发
作者：
Shigeru Sugiyama;Noriko Shimizu;Keisuke Kakinouchi;Osamu Hiraoka;Hiroyoshi Matsumura;Hiroshi Y. Yoshikawa;Yoshinori Takahashi;Mihoko Maruyama;Masashi Yoshimura;Hiroaki Adachi;Kazufumi Takano;Satoshi Murakami;Tsuyoshi Inoue;Michio Murata;and
中子大分子晶体学可以确定蛋白质中氢原子的位置，从而阐明蛋白质的结构和功能以及酶的作用机制。它需要非常大的晶体来弥补可用中子束的低通量。已经开发了一些技术来获得大的蛋白质晶体。然而，晶体的处理仍然存在问题。在这里，我们提出了一种新的结晶技术，结合高强度水凝胶和接种方法与脂肪酸结合蛋白和溶菌酶。这种新的方法能够安全地将种子引入到预平衡的蛋白质溶液中，而不会造成损坏。因此，我们可以生长大的单晶体而不形成孪晶和多晶。这项技术有望扩大重要蛋白质的范围，并可能导致高分辨率X射线和中子晶体学的自动化系统。
Neutron macromolecule crystallography can determine the positions of hydrogen atoms in proteins, thereby clarifying the protein structure and function as well as enzyme mechanisms. It requires very large crystals to make up for the low flux of available neutron beams. Some techniques have been developed to obtain large protein crystals. However, problems remain with the handling of the crystal. Here we propose a novel crystallization technique that combines high-strength hydrogel and seeding methods with fatty-acid binding protein and lysozyme. This novel approach enables safe introduction of seeds into pre-equilibrated protein solutions without damage. Consequently, we can grow large single crystals without forming twinned crystals and polycrystals. This technique is expected to expand the scope of important proteins and may lead to an automated system for high-resolution X-ray and neutron crystallography.