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The active site protonation states of perdeuterated Toho‐1 β‐lactamase determined by neutron diffraction support a role for Glu166 as the general base in acylation
Author(s) -
Tomanicek Stephen J.,
Wang Kathy K.,
Weiss Kevin L.,
Blakeley Matthew P.,
Cooper Jonathan,
Chen Yu,
Coates Leighton
Publication year - 2011
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2010.12.017
Subject(s) - protonation , active site , neutron diffraction , crystallography , acylation , hydrogen bond , chemistry , deuterium , stereochemistry , enzyme , crystal structure , molecule , physics , catalysis , biochemistry , organic chemistry , atomic physics , ion
Room temperature neutron diffraction data of the fully perdeuterated Toho‐1 R274N/R276N double mutant β‐lactamase in the apo form were used to visualize deuterium atoms within the active site of the enzyme. This perdeuterated neutron structure of the Toho‐1 R274N/R276N reveals the clearest picture yet of the ground‐state active site protonation states and the complete hydrogen‐bonding network in a β‐lactamase enzyme. The ground‐state active site protonation states detailed in this neutron diffraction study are consistent with previous high‐resolution X‐ray studies that support the role of Glu166 as the general base during the acylation reaction in the class A β‐lactamase reaction pathway.