Towards a full quantitative description of yeast metabolism: a systematic approach for estimating the kinetic parameters of isoenzymes under in vivo like conditions

Hanan L Messiha, Naglis Malys, Kathleen M Carroll

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

In order to produce a full quantitative description of yeast metabolism, a number of kinetic parameters of enzymes that are important for energy metabolism must be determined experimentally. We aim to determine the prospective in vivo kinetic properties of a range of yeast-purified isoenzymes that are important in energy metabolism, with respect to the concentration of their substrates and products. This endeavor forms part of our systems biology pipeline to facilitate the production of bottom-up models of metabolism. Within this workflow, we implement an infrastructure for medium- to high-throughput determination of the kinetic properties of purified isoenzymes in in vivo like conditions. This includes the use of the KineticsWizard software for data capture and analysis. The captured experimental data are analyzed by the software and subsequently stored in appropriate repositories (MeMo-RK and SABIO-RK). While we focus initially on glycolysis in Saccharomyces cerevisiae, our methodology is generic and can be widely applied to the study of other enzymes and pathways in yeast and other organisms.

Original languageEnglish
Title of host publicationMethods in Enzymology
Subtitle of host publicationMethods in Systems Biology
EditorsDaniel Jameson, Malkhey Verma, Hans V. Westerhoff
Place of PublicationSan Diego
PublisherAcademic Press
Chapter12
Pages215-231
Number of pages17
ISBN (Print)9780123851185
DOIs
Publication statusPublished - 12 Nov 2011

Publication series

NameMethods in Enzymology
PublisherAcademic Press
Volume500
ISSN (Print)0076-6879

Keywords

  • yeast metabolism
  • kinetic parameters
  • enzyme assays
  • glycolytic isoenzyme
  • KineticsWizard
  • pyruvate decarboxylase

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