Structural and kinetic properties of NADP-malic enzyme from the inducible crassulacean acid metabolism plant mesembryanthemum crystallinum l.

Kazuyuki Saitou, Waichi Agata, Masae Asakura, Fumitake Kubota

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Abstract

NADP-malic enzyme (EC 1.1.1.40), which is involved in Crassulacean acid metabolism (CAM), was purified to electrophoretic homogeneity from the leaves of the inducible CAM plant Mesembryanthemum crystallinum. The NADP-malic enzyme, which was purified 1,146-fold, has a specific activity of 68.8 μmol (mg protein)-1 min-1. The molecular weight of the subunits of the enzyme was 64 kDa. The native molecular weight of the enzyme was determined by gel-filtration to be 390 kDa, indicating that the purified NADP-malic enzyme is a hexamer of identical subunits. The optimal pH for activity of the enzyme was around 7.2. Double-reciprocal plots of the enzymatic activity as a function of the concentration of L-malate yielded straight lines both at pH 7.2 and at pH 7.8 and did not reveal any evidence for cooperativity of binding of L-malate. The Km value for L-malate was 0.35 mM. Hill plots of the activity as a function of the concentration of NADP+ indicated positive cooperativity in the binding of NADP+ to the enzyme with a Hill coefficient (nH) of 2.0. An S0.5 value (the concentration giving half-maximal activity) of 9.9 μM for NADP+ was obtained. Oxaloacetate inhibited the activity of the NADP-malic enzyme. Effects of succinate and NaHCO3 on the activity of NADP-malic enzyme were small.

Original languageEnglish
Pages (from-to)595-600
Number of pages6
JournalPlant and Cell Physiology
Volume33
Issue number5
Publication statusPublished - Jul 1 1992

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malate dehydrogenase (oxaloacetate-decarboxylating) (NADP+)
Mesembryanthemum
Mesembryanthemum crystallinum
Crassulacean acid metabolism
malic enzyme
kinetics
malates
Acids
NADP
Enzymes
Molecular Weight
malate dehydrogenase (oxaloacetate-decarboxylating) (NADP)
molecular weight
Oxaloacetic Acid
Succinic Acid
protein subunits
succinic acid
enzymes
Gel Chromatography
gels

All Science Journal Classification (ASJC) codes

  • Medicine(all)
  • Physiology
  • Plant Science
  • Cell Biology

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Structural and kinetic properties of NADP-malic enzyme from the inducible crassulacean acid metabolism plant mesembryanthemum crystallinum l. / Saitou, Kazuyuki; Agata, Waichi; Asakura, Masae; Kubota, Fumitake.

In: Plant and Cell Physiology, Vol. 33, No. 5, 01.07.1992, p. 595-600.

Research output: Contribution to journalArticle

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