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Title: The active site protonation states of perdeuterated Toho-1 beta-lactamase determined by neutron diffraction support a role for Glu166 as the general base in acylation

Journal Article · · FEBS Letters
 [1];  [1]; ORCiD logo [1];  [2];  [3]; ORCiD logo [1];  [4]
  1. ORNL
  2. Institut Laue-Langevin (ILL)
  3. University of Southampton, UK
  4. University of South Florida
+ Show Author Affiliations

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.

Research Organization:
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
Sponsoring Organization:
USDOE
DOE Contract Number:
AC05-00OR22725
OSTI ID:
1037160
Journal Information:
FEBS Letters, Vol. 585, Issue 21168411; ISSN 0014-5793
Country of Publication:
United States
Language:
English

References (26)

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Site-Directed Mutagenesis of Glutamate-166 in .beta.-Lactamase Leads to a Branched Path Mechanism journal June 1994
Growing Group of Extended-Spectrum  -Lactamases: the CTX-M Enzymes journal December 2003
Identification of Glu166 as the General Base in the Acylation Reaction of Class A β-Lactamases through QM/MM Modeling journal August 2003
An Ultrahigh Resolution Structure of TEM-1 β-Lactamase Suggests a Role for Glu166 as the General Base in Acylation journal May 2002
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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
60 APPLIED LIFE SCIENCES
ACYLATION
ATOMS
DEUTERIUM
ENZYMES
MUTANTS
NEUTRON DIFFRACTION