Properties of purified methylmalonate semialdehyde dehydrogenase of Pseudomonas aeruginosa.

Abstract Methylmalonate semialdehyde dehydrogenase, an inducible enzyme from Pseudomonas aeruginosa which catalyzes the oxidation of methylmalonate semialdehyde to propionyl coenzyme A, was purified to the extent that a preparation was obtained which was homogenous by the criterion of disc gel electrophoresis. The molecular weight of the native enzyme was estimated to be 132,000 by gel filtration. Methylmalonate semialdehyde dehydrogenase dissociated into subunits in urea or sodium dodecyl sulfate solutions. The molecular weight of the subunit was estimated at between 58,000 and 69,000 by the techniques of gel electrophoresis and gel filtration in sodium dodecyl sulfate, respectively, which means that the native enzyme is composed of at least two subunits. The purified enzyme catalyzed the reduction of NAD with methylmalonate semialdehyde and propionaldehyde. In the presence of CoA, propionyl-CoA was a product of methylmalonate semialdehyde oxidation. The enzyme was active when mercaptoethanol replaced CoA and an active propionyl derivative was formed then as well. The stoichiometry between NADH and propionyl-CoA was 1:1. 3-Hydroxyisobutyrate dehydrogenase and 3-hydroxyisobutyrate-1-14C were used to generate methylmalonate semialdehyde-1-14C, and radioactive carbon dioxide was produced when methylmalonate semialdehyde dehydrogenase was present in the reaction mixture. The Michaelis constants for methylmalonate semialdehyde, NAD, CoA, mercaptoethanol, and propionaldehyde were determined. The proposal is made that the physiological role of methylmalonate semialdehyde dehydrogenase in cell metabolism is to catalyze the oxidation of methylmalonate semialdehyde to propionyl-CoA in the catabolism of valine.

• Publication year

  1970

• Venue

  Journal of Biological Chemistry

• Publication date

  1970-04-10

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1016/S0021-9258(19)77166-XPMID 4314598
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• In the presence of CoA, the enzyme formed propionyl-CoA with a 1:1 stoichiometry relative to NADH.

  Confidence 0.94

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) review
• The enzyme is proposed to function in valine catabolism by oxidizing methylmalonate semialdehyde to propionyl-CoA.

  Confidence 0.95

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) review
• Methylmalonate semialdehyde dehydrogenase dissociated into subunits in urea or sodium dodecyl sulfate, indicating that the native enzyme contains at least two subunits.

  Confidence 0.97

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) review
• Methylmalonate semialdehyde dehydrogenase from Pseudomonas aeruginosa was purified to homogeneity by disc gel electrophoresis, and its native molecular weight was estimated at about 132,000 by gel filtration.

  Confidence 0.98

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) review

• disc gel electrophoresis

  method, analytical technique

  A protein-separation method used here to assess whether the enzyme preparation was homogeneous.

  Aliases: disc PAGE

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) review
• gel filtration

  method, analytical technique

  A size-based separation method used here to estimate the native molecular weight of the enzyme.

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) review
• methylmalonate semialdehyde dehydrogenase

  enzyme, protein

  An inducible dehydrogenase enzyme purified from Pseudomonas aeruginosa that acts on methylmalonate semialdehyde.

  Aliases: MSADH

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) review
• propionyl-coa

  metabolite, product

  A coenzyme A thioester product formed during oxidation of methylmalonate semialdehyde in the assay.

  Aliases: propionyl coenzyme A

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) review
• pseudomonas aeruginosa

  organism, bacterium

  A bacterial species used as the source organism for the purified enzyme.

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) review
• sodium dodecyl sulfate

  reagent, detergent

  A detergent used here to dissociate the enzyme into subunits and estimate subunit size.

  Aliases: SDS

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) review
• urea

  reagent, denaturant

  A denaturing reagent used here to promote dissociation of the enzyme into subunits.

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) review
• valine catabolism

  pathway, metabolic process

  The metabolic pathway proposed as the physiological context for the enzyme's activity.

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) review

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