Kinetic behavior of the Arabidopsis thaliana leaf formate dehydrogenase is thermally sensitive

Renee D. Baack, John Markwell, Patricia L. Herman, John Osterman

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Two previous kinetic studies on the Arabidopsis thaliana leaf NAD-dependent formate dehydrogenase (EC 1.2.1.2) have demonstrated two very different sets of Km values for the formate and NAD+ substrates. We examined the kinetics of the enzyme partially purified from a leaf extract by gel-filtration desalting and chromatography on DEAE-cellulose, as well as by isolation of a mitochondria-enriched fraction obtained by differential centrifugation. Both of these methods produce a formate dehydrogenase enzyme with the higher Km values of approximately 10 mmol/L formate and 75 μmol/L NAD+. The kinetic properties of the Arabidopsis formate dehydrogenase expressed to high levels in transgenic tobacco plants were also those of the high Km form. The high Km form of the enzyme converted to a low Km form by heating for 5 minutes at 60°C. An Arrhenius plot of the activity during the heating process was linear, indicating that the heating did not cause alterations in either the active site or the thermal dependence of the catalytic reaction. We conclude that the native form of the formate dehydrogenase probably resembles the form with the higher Km values. Heating seemingly converts this native enzyme to the molten globule state and cooling results in formation of a non-native structure with altered kinetic properties.

Original languageEnglish (US)
Pages (from-to)445-450
Number of pages6
JournalJournal of Plant Physiology
Volume160
Issue number5
DOIs
StatePublished - May 1 2003

Fingerprint

Formate Dehydrogenases
formic acid
Arabidopsis
Heating
Arabidopsis thaliana
heat
kinetics
Enzymes
NAD
formates
leaves
DEAE-Cellulose
enzymes
Genetically Modified Plants
Centrifugation
enzyme kinetics
Tobacco
Gel Chromatography
Catalytic Domain
Mitochondria

Keywords

  • Arabidopsis thaliana
  • Formate
  • Formate dehydrogenase
  • Native enzyme

ASJC Scopus subject areas

  • Physiology
  • Agronomy and Crop Science
  • Plant Science

Cite this

Kinetic behavior of the Arabidopsis thaliana leaf formate dehydrogenase is thermally sensitive. / Baack, Renee D.; Markwell, John; Herman, Patricia L.; Osterman, John.

In: Journal of Plant Physiology, Vol. 160, No. 5, 01.05.2003, p. 445-450.

Research output: Contribution to journalArticle

Baack, Renee D. ; Markwell, John ; Herman, Patricia L. ; Osterman, John. / Kinetic behavior of the Arabidopsis thaliana leaf formate dehydrogenase is thermally sensitive. In: Journal of Plant Physiology. 2003 ; Vol. 160, No. 5. pp. 445-450.
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