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PMID: 20498365 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Glutathione synthesis and turnover in the human erythrocyte: alignment of a model based on detailed enzyme kinetics with experimental data.

The Journal of biological chemistry ·Vol. 285 ·No. 31 ·2010-07-30 ·Pages 23557-67

Raftos JE, Whillier S, Kuchel PW

Abstract

The erythrocyte is exposed to reactive oxygen species in the circulation and also to those produced by autoxidation of hemoglobin. Consequently, erythrocytes depend on protection by the antioxidant glutathione. Mathematical models based on realistic kinetic data have provided valuable insights into the regulation of biochemical pathways within the erythrocyte but none have satisfactorily accounted for glutathione metabolism. In the current model, rate equations were derived for the enzyme-catalyzed reactions, and for each equation the nonlinear algebraic relationship between the steady-state kinetic parameters and the unitary rate constants was derived. The model also includes the transport processes that supply the amino acid constituents of glutathione and the export of oxidized glutathione. Values of the kinetic parameters for the individual reactions were measured predominately using isolated enzymes under conditions that differed from the intracellular environment. By comparing the experimental and simulated results, the values of the enzyme-kinetic parameters of the model were refined to yield conformity between model simulations and experimental data. Model output accurately represented the steady-state concentrations of metabolites in erythrocytes suspended in plasma and the changing glutathione concentrations in whole and hemolyzed erythrocytes under specific experimental conditions. Analysis indicated that feedback inhibition of gamma-glutamate-cysteine ligase by glutathione had a limited effect on steady-state glutathione concentrations and was not sufficiently potent to return glutathione concentrations to normal levels in erythrocytes exposed to sustained increases in oxidative load.

MeSH Terms
Adult Alanine Transaminase/chemistry Catalysis Erythrocytes/metabolism Gene Expression Regulation Glutamate-Cysteine Ligase/metabolism Glutaminase/chemistry Glutathione/metabolism Humans Kinetics Middle Aged Models, Biological Models, Theoretical Oxidative Stress Oxygen/chemistry
Chemicals
Alanine Transaminase Glutaminase Glutamate-Cysteine Ligase Glutathione Oxygen
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Raftos Julia E
Department of Biological Sciences, Macquarie University, Sydney, New South Wales 2109, Australia. [email protected]
Whillier Stephney
Kuchel Philip W
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
1083-351X
DOI
10.1074/jbc.M109.067017
Email
[email protected]
Published
2010-07-30
Epub
2010-00-24
Pages
23557-67
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC2911318
Subset
IM
External Links
PubMed source DOI: 10.1074/jbc.M109.067017 PMC Full Text View journal details
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