Shared promiscuous activities and evolutionary features in various members of the amidohydrolase superfamily. Export

@article{citeulike:399761, abstract = {The amidohydrolase superfamily comprises hundreds of hydrolytic enzymes of the (beta/alpha)8 barrel fold with mono- or binuclear active-site metal centers, and a diverse spectrum of substrates and reactions. Promiscuous activities, or cross-reactivities, between different members of the same superfamily may provide important hints regarding evolutionary and mechanistic relationships. We examined three members: dihydroorotase (DHO), phosphotriesterase (PTE), and PTE-homology protein (PHP). Of particular interest are PTE, which is thought to have evolved within the last several decades, and PHP, an amidohydrolase superfamily member of unknown function, and the closest known homologue of PTE. We found a diverse and partially overlapping pattern of promiscuous activities in these enzymes, including a significant lactonase activity in PTE, esterase activities in both PTE and PHP, and a weak PTE activity in DHO. Directed evolution was applied to improve the promiscuous esterase activities of PTE and PHP. Remarkably, the most recurrent mutation increasing esterase activity in PTE, or PHP, maps to the same location in their superposed 3D structures. The evolved variants also exhibit newly acquired promiscuous activities that were not selected for, including very weak, yet measurable, paraoxonase activity in PHP. Our results illustrate the mechanistic, structural, and evolutionary links between these enzymes, and highlight the importance of studying laboratory evolution intermediates that might resemble node intermediates along the evolutionary pathways leading to the divergence of enzyme superfamilies.}, address = {Department of Biological Chemistry, The Weizmann Institute of Science, 76100 Rehovot, Israel.}, author = {Roodveldt, C. and Tawfik, D. S.}, citeulike-article-id = {399761}, citeulike-linkout-0 = {http://dx.doi.org/10.1021/bi051021e}, citeulike-linkout-1 = {http://pubs.acs.org/doi/abs/10.1021/bi051021e}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/16171387}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=16171387}, day = {27}, doi = {10.1021/bi051021e}, issn = {0006-2960}, journal = {Biochemistry}, keywords = {enzyme, mechanism, plasticity, struct\_func}, month = {September}, number = {38}, pages = {12728--12736}, posted-at = {2005-11-18 12:08:42}, priority = {4}, title = {Shared promiscuous activities and evolutionary features in various members of the amidohydrolase superfamily.}, url = {http://dx.doi.org/10.1021/bi051021e}, volume = {44}, year = {2005} }

TY - JOUR ID - citeulike:399761 L3 - citeulike-article-id:399761 N2 - The amidohydrolase superfamily comprises hundreds of hydrolytic enzymes of the (beta/alpha)8 barrel fold with mono- or binuclear active-site metal centers, and a diverse spectrum of substrates and reactions. Promiscuous activities, or cross-reactivities, between different members of the same superfamily may provide important hints regarding evolutionary and mechanistic relationships. We examined three members: dihydroorotase (DHO), phosphotriesterase (PTE), and PTE-homology protein (PHP). Of particular interest are PTE, which is thought to have evolved within the last several decades, and PHP, an amidohydrolase superfamily member of unknown function, and the closest known homologue of PTE. We found a diverse and partially overlapping pattern of promiscuous activities in these enzymes, including a significant lactonase activity in PTE, esterase activities in both PTE and PHP, and a weak PTE activity in DHO. Directed evolution was applied to improve the promiscuous esterase activities of PTE and PHP. Remarkably, the most recurrent mutation increasing esterase activity in PTE, or PHP, maps to the same location in their superposed 3D structures. The evolved variants also exhibit newly acquired promiscuous activities that were not selected for, including very weak, yet measurable, paraoxonase activity in PHP. Our results illustrate the mechanistic, structural, and evolutionary links between these enzymes, and highlight the importance of studying laboratory evolution intermediates that might resemble node intermediates along the evolutionary pathways leading to the divergence of enzyme superfamilies. IS - 38 JF - Biochemistry SN - 0006-2960 AD - Department of Biological Chemistry, The Weizmann Institute of Science, 76100 Rehovot, Israel. EP - 12736 TI - Shared promiscuous activities and evolutionary features in various members of the amidohydrolase superfamily. VL - 44 SP - 12728 KW - enzyme KW - mechanism KW - plasticity KW - struct_func AU - Roodveldt, C AU - Tawfik, DS PY - 2005/09/27/ UR - http://dx.doi.org/10.1021/bi051021e ER -