Bovine liver formaldehyde dehydrogenase. Kinetic and molecular properties

Tayebeh Pourmotabbed, M. J. Shih, D. J. Creighton

Research output: Contribution to journalArticle

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Abstract

The dimeric formaldehyde dehydrogenase from bovine liver has been resolved into three nearly homogeneous enzyme forms by the successive use of ion-exchange, affinity, and ampholine (chromatofocusing) chromatography. The different enzyme species were isolated in the approximate proportions 3:2:1, having pI values of 6.5, 6.2, and 6.0, respectively. The subunit molecular weights of the three forms are all similar (M(r) ≃ 41,000), on the basis of sodium dodecyl sulfate-gel electrophoresis. The enzyme species appear to arise from covalent differences unrelated either to partial proteolysis during isolation or to differential sialization of homodimeric protein. Human liver contains a single major form and two minor forms of formaldehyde dehydrogenase having pI values very similar to those found for the bovine liver enzyme. The macroscopic kinetic constants (V, V/K) for the three forms of the dehydrogenase from bovine liver are all similar in magnitude, using NADH and S-hydroxymethylglutathione as substrates. The isotope-sensitive hydride transfer step is not significantly rate-limiting during catalysis by any of the forms, as evidenced by the near-unity primary deuterium isotope effects on both V and V/K(s) (for S-hydroxymethylglutathione); catalysis may be limited by the rate of dissociation of at least one (and possibly both) of the product molecules. In support of rate-limiting dissociation of NAD+ in the normal reaction, V increases by ~ 22-fold and isotope effects of ~ 1.4 are observed on both V and V/K(s), using the coenzyme analog 3-acetylpyridine adenine dinucleotide. Product dissociation from the active site appears to be accelerated by the presence of dilute denaturing agents, perhaps indicative of a rate-limiting conformational transition associated with product release.

Original languageEnglish (US)
Pages (from-to)17384-17388
Number of pages5
JournalJournal of Biological Chemistry
Volume264
Issue number29
StatePublished - Jan 1 1989
Externally publishedYes

Fingerprint

glutathione-independent formaldehyde dehydrogenase
Liver
Isotopes
Kinetics
Enzymes
Catalysis
NAD
Proteolysis
Deuterium
Ion Exchange
Coenzymes
Chromatography
Electrophoresis
Hydrides
Sodium Dodecyl Sulfate
Catalytic Domain
Ion exchange
Oxidoreductases
Molecular Weight
Gels

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Bovine liver formaldehyde dehydrogenase. Kinetic and molecular properties. / Pourmotabbed, Tayebeh; Shih, M. J.; Creighton, D. J.

In: Journal of Biological Chemistry, Vol. 264, No. 29, 01.01.1989, p. 17384-17388.

Research output: Contribution to journalArticle

Pourmotabbed, Tayebeh ; Shih, M. J. ; Creighton, D. J. / Bovine liver formaldehyde dehydrogenase. Kinetic and molecular properties. In: Journal of Biological Chemistry. 1989 ; Vol. 264, No. 29. pp. 17384-17388.
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