Tsuda Tomohiro, Nihira Takanori, Chiku Kazuhiro, Suzuki Erika, Arakawa Takatoshi, Nishimoto Mamoru, Kitaoka Motomitsu, Nakai Hiroyuki, Fushinobu Shinya
Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Faculty of Agriculture, Niigata University, Niigata 950-2181, Japan.
FEBS Lett. 2015 Dec 21;589(24 Pt B):3816-21. doi: 10.1016/j.febslet.2015.11.034. Epub 2015 Nov 26.
Glycoside hydrolase family 130 consists of phosphorylases and hydrolases for β-mannosides. Here, we characterized β-1,2-mannobiose phosphorylase from Listeria innocua (Lin0857) and determined its crystal structures complexed with β-1,2-linked mannooligosaccharides. β-1,2-Mannotriose was bound in a U-shape, interacting with a phosphate analog at both ends. Lin0857 has a unique dimer structure connected by a loop, and a significant open-close loop displacement was observed for substrate entry. A long loop, which is exclusively present in Lin0857, covers the active site to limit the pocket size. A structural basis for substrate recognition and phosphorolysis was provided.
糖苷水解酶家族130由β-甘露糖苷的磷酸化酶和水解酶组成。在此，我们对无害李斯特菌的β-1,2-甘露二糖磷酸化酶（Lin0857）进行了表征，并确定了其与β-1,2-连接的甘露寡糖复合的晶体结构。β-1,2-甘露三糖呈U形结合，在两端与磷酸类似物相互作用。Lin0857具有通过一个环连接的独特二聚体结构，并且观察到底物进入时环有明显的开闭位移。一个仅存在于Lin0857中的长环覆盖活性位点以限制口袋大小。提供了底物识别和磷酸解的结构基础。
