Protein engineering of a human enzyme that hydrolyzes V and G nerve agents

Design, construction and characterization

Clarence A. Broomfield, Oksana Lockridge, Charles B. Millard

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Because of deficiencies in the present treatments for organophosphorus anticholinesterase poisoning, we are attempting to develop a catalytic scavenger that can be administered as prophylactic protection. Currently known enzymes are inadequate for this purpose because they have weak binding and slow turnover, so we are trying to make an appropriate enzyme by protein engineering techniques. One butyrylcholinesterase mutant, G117H, has the desired type of activity but reacts much too slowly. This communication describes an attempt to determine the reason for the slow reaction so that a more efficient enzyme might be designed. The results indicate that the mutation at residue 117 has resulted in a distortion of the transition state of the reaction of organophosphorus compounds with the active site serine. This information will be used to develop other mutants that avoid transition state stabilization sites. Copyright (C) 1999 Elsevier Science Ireland Ltd.

Original languageEnglish (US)
Pages (from-to)413-418
Number of pages6
JournalChemico-Biological Interactions
Volume119-120
DOIs
StatePublished - May 14 1999

Fingerprint

Protein Engineering
Enzymes
Organophosphate Poisoning
Organophosphorus Compounds
Butyrylcholinesterase
Proteins
Cholinesterase Inhibitors
Serine
Catalytic Domain
Stabilization
Mutation
Communication
Nerve Agents

Keywords

  • Butyrylcholinesterase
  • Enzyme engineering
  • OPA hydrolase
  • Organophosphorus anticholinesterases
  • Site-directed mutagenesis
  • Transition state

ASJC Scopus subject areas

  • Toxicology

Cite this

Protein engineering of a human enzyme that hydrolyzes V and G nerve agents : Design, construction and characterization. / Broomfield, Clarence A.; Lockridge, Oksana; Millard, Charles B.

In: Chemico-Biological Interactions, Vol. 119-120, 14.05.1999, p. 413-418.

Research output: Contribution to journalArticle

@article{2c9e4b54f34c4a4eb96731aaf7097075,
title = "Protein engineering of a human enzyme that hydrolyzes V and G nerve agents: Design, construction and characterization",
abstract = "Because of deficiencies in the present treatments for organophosphorus anticholinesterase poisoning, we are attempting to develop a catalytic scavenger that can be administered as prophylactic protection. Currently known enzymes are inadequate for this purpose because they have weak binding and slow turnover, so we are trying to make an appropriate enzyme by protein engineering techniques. One butyrylcholinesterase mutant, G117H, has the desired type of activity but reacts much too slowly. This communication describes an attempt to determine the reason for the slow reaction so that a more efficient enzyme might be designed. The results indicate that the mutation at residue 117 has resulted in a distortion of the transition state of the reaction of organophosphorus compounds with the active site serine. This information will be used to develop other mutants that avoid transition state stabilization sites. Copyright (C) 1999 Elsevier Science Ireland Ltd.",
keywords = "Butyrylcholinesterase, Enzyme engineering, OPA hydrolase, Organophosphorus anticholinesterases, Site-directed mutagenesis, Transition state",
author = "Broomfield, {Clarence A.} and Oksana Lockridge and Millard, {Charles B.}",
year = "1999",
month = "5",
day = "14",
doi = "10.1016/S0009-2797(99)00053-8",
language = "English (US)",
volume = "119-120",
pages = "413--418",
journal = "Chemico-Biological Interactions",
issn = "0009-2797",
publisher = "Elsevier Ireland Ltd",

}

TY - JOUR

T1 - Protein engineering of a human enzyme that hydrolyzes V and G nerve agents

T2 - Design, construction and characterization

AU - Broomfield, Clarence A.

AU - Lockridge, Oksana

AU - Millard, Charles B.

PY - 1999/5/14

Y1 - 1999/5/14

N2 - Because of deficiencies in the present treatments for organophosphorus anticholinesterase poisoning, we are attempting to develop a catalytic scavenger that can be administered as prophylactic protection. Currently known enzymes are inadequate for this purpose because they have weak binding and slow turnover, so we are trying to make an appropriate enzyme by protein engineering techniques. One butyrylcholinesterase mutant, G117H, has the desired type of activity but reacts much too slowly. This communication describes an attempt to determine the reason for the slow reaction so that a more efficient enzyme might be designed. The results indicate that the mutation at residue 117 has resulted in a distortion of the transition state of the reaction of organophosphorus compounds with the active site serine. This information will be used to develop other mutants that avoid transition state stabilization sites. Copyright (C) 1999 Elsevier Science Ireland Ltd.

AB - Because of deficiencies in the present treatments for organophosphorus anticholinesterase poisoning, we are attempting to develop a catalytic scavenger that can be administered as prophylactic protection. Currently known enzymes are inadequate for this purpose because they have weak binding and slow turnover, so we are trying to make an appropriate enzyme by protein engineering techniques. One butyrylcholinesterase mutant, G117H, has the desired type of activity but reacts much too slowly. This communication describes an attempt to determine the reason for the slow reaction so that a more efficient enzyme might be designed. The results indicate that the mutation at residue 117 has resulted in a distortion of the transition state of the reaction of organophosphorus compounds with the active site serine. This information will be used to develop other mutants that avoid transition state stabilization sites. Copyright (C) 1999 Elsevier Science Ireland Ltd.

KW - Butyrylcholinesterase

KW - Enzyme engineering

KW - OPA hydrolase

KW - Organophosphorus anticholinesterases

KW - Site-directed mutagenesis

KW - Transition state

UR - http://www.scopus.com/inward/record.url?scp=0033001689&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033001689&partnerID=8YFLogxK

U2 - 10.1016/S0009-2797(99)00053-8

DO - 10.1016/S0009-2797(99)00053-8

M3 - Article

VL - 119-120

SP - 413

EP - 418

JO - Chemico-Biological Interactions

JF - Chemico-Biological Interactions

SN - 0009-2797

ER -