Decarbamoylation of acetylcholinesterases is markedly slowed as carbamoyl groups increase in size

Kunisi S. Venkatasubban, Joseph L. Johnson, Jamie L. Thomas, Abdul Fauq, Bernadette Cusack, Terrone L. Rosenberry

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

Carbamates are esters of substituted carbamic acids that react with acetylcholinesterase (AChE) by initially transferring the carbamoyl group to a serine residue in the enzyme active site accompanied by loss of the carbamate leaving group followed by hydrolysis of the carbamoyl enzyme. This hydrolysis, or decarbamoylation, is relatively slow, and half-lives of carbamoylated AChEs range from 4 min to more than 30 days. Therefore, carbamates are effective AChE inhibitors that have been developed as insecticides and as therapeutic agents. We show here, in contrast to a previous report, that decarbamoylation rate constants are independent of the leaving group for a series of carbamates with the same carbamoyl group. When the alkyl substituents on the carbamoyl group increased in size from N-monomethyl- to N,N-dimethyl-, N-ethyl-N-methyl-, or N,N-diethyl-, the decarbamoylation rate constants decreased by 4-, 70-, and 800-fold, respectively. We suggest that this relationship arises as a result of active site distortion, particularly in the acyl pocket of the active site. Furthermore, solvent deuterium oxide isotope effects for decarbamoylation decreased from 2.8 for N-monomethylcarbamoyl AChE to 1.1 for N,N-diethylcarbamoyl AChE, indicating a shift in the rate-limiting step from general acid-base catalysis to a likely conformational change in the distorted active site.

Original languageEnglish (US)
Pages (from-to)67-74
Number of pages8
JournalArchives of Biochemistry and Biophysics
Volume655
DOIs
StatePublished - Oct 1 2018

Bibliographical note

Funding Information:
This work was supported by grant NS-16577 from the National Institutes of Health , by grant DAMD 17-98-2-8019 from the United States Army Medical Research Acquisition Activity , by grants from the Muscular Dystrophy Association of America to T.L.R , and by NIH NRSA fellowship NS-41828 to J.L.J.

Keywords

  • Acetylcholinesterase
  • Alzheimer's disease
  • Decarbamoylation

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