structures of helicobacter pylori shikimate kinase reveal a selective inhibitor-induced-fit mechanism幽门螺杆菌shikimate激酶结构揭示了选择性inhibitor-induced-fit机制.pdf

Structures of Helicobacter pylori Shikimate Kinase Reveal a Selective Inhibitor-Induced-Fit Mechanism 1,2. 3. 1,2 3 1 Wen-Chi Cheng , Yen-Fu Chen , Hung-Jung Wang , Kai-Cheng Hsu , Shuang-Chih Lin , Tzu-Jung Chen1, Jinn-Moon Yang3,4*, Wen-Ching Wang1,2* 1 Institute of Molecular and Cellular Biology and Department of Life Sciences, National Tsing Hua University, Hsinchu, Taiwan, 2 Biomedical Science and Engineering Center, National Tsing Hua University, Hsinchu, Taiwan, 3 Institute of Bioinformatics and Systems Biology, National Chiao Tung University, Hsinchu, Taiwan, 4 Department of Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan Abstract Shikimate kinase (SK), which catalyzes the specific phosphorylation of the 3-hydroxyl group of shikimic acid in the presence of ATP, is the enzyme in the fifth step of the shikimate pathway for biosynthesis of aromatic amino acids. This pathway is present in bacteria, fungi, and plants but absent in mammals and therefore represents an attractive target pathway for the development of new antimicrobial agents, herbicides, and antiparasitic agents. Here we investigated the detailed structure– activity relationship of SK from Helicobacter pylori (HpSK). Site-directed mutagenesis and isothermal titration calorimetry studies revealed critical conserved residues (D33, F48, R57, R116, and R132) that interact with shikimate and are therefore involved in catalysis. Crystal structures of HpSK?SO4, R57A, and HpSKNshikimate-3-phosphateNADP show a characteristic three-layer architecture and a conformationally elastic region consisting of F48, R57, R116, and R13