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

Stephen J. Tomanicek; Kathy K. Wang; Kevin L. Weiss; Matthew P. Blakeley; Jonathan Cooper; Yu Chen; Leighton Coates

doi:10.1016/j.febslet.2010.12.017

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

Stephen J. Tomanicek, Kathy K. Wang, Kevin L. Weiss, Matthew P. Blakeley, Jonathan Cooper, Yu Chen, Leighton Coates

Research output: Contribution to journal › Article › peer-review

34 Scopus citations

Abstract

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.

Original language	English
Pages (from-to)	364-368
Number of pages	5
Journal	FEBS Letters
Volume	585
Issue number	2
DOIs	https://doi.org/10.1016/j.febslet.2010.12.017
State	Published - Jan 21 2011

Funding

This research was sponsored by the Laboratory Directed Research and Development Program of Oak Ridge National Laboratory (ORNL) , managed by UT-Battelle LLC for the US Department of Energy under Contract No. DE-AC05-00OR22725.

Funders	Funder number
U.S. Department of Energy	DE-AC05-00OR22725
Oak Ridge National Laboratory
UT-Battelle

Keywords

CTX-M-type ESBLs
Extended-spectrum β-lactamases
Neutron diffraction
Perdeuterated neutron structure
Toho-1
β-Lactamase

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10.1016/j.febslet.2010.12.017

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title = "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",

abstract = "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.",

keywords = "CTX-M-type ESBLs, Extended-spectrum β-lactamases, Neutron diffraction, Perdeuterated neutron structure, Toho-1, β-Lactamase",

author = "Tomanicek, {Stephen J.} and Wang, {Kathy K.} and Weiss, {Kevin L.} and Blakeley, {Matthew P.} and Jonathan Cooper and Yu Chen and Leighton Coates",

year = "2011",

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doi = "10.1016/j.febslet.2010.12.017",

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T1 - 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

AU - Tomanicek, Stephen J.

AU - Wang, Kathy K.

AU - Weiss, Kevin L.

AU - Blakeley, Matthew P.

AU - Cooper, Jonathan

AU - Chen, Yu

AU - Coates, Leighton

PY - 2011/1/21

Y1 - 2011/1/21

N2 - 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.

AB - 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.

KW - CTX-M-type ESBLs

KW - Extended-spectrum β-lactamases

KW - Neutron diffraction

KW - Perdeuterated neutron structure

KW - Toho-1

KW - β-Lactamase

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DO - 10.1016/j.febslet.2010.12.017

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AN - SCOPUS:78651374505

SN - 0014-5793

VL - 585

SP - 364

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IS - 2

ER -

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

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Keywords

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