The fate of fluoropyruvate as an alternative substrate and inactivator of the pyruvate dehydrogenase complex of Escherichia coli
Thesis/Dissertation
·
OSTI ID:5564220
The pyruvate dehydrogenase component (E{sub 1}) of the pyruvate dehydrogenase complex of Escherichia coli catalyzes the decomposition of fluoropyruvate to CO{sub 2}, fluoride anion, and acetate. Acetyl thiamin pyrophosphate (acetyl-TPP) is an intermediate in this reaction. Incubation of the pyruvate dehydrogenase complex with ({sup 14}C)fluoropyruvate, TPP, CoASH, and either NADH or pyruvate as reducing agent leads to the formation of ({sup 14}C) acetylCoA. In this reaction the acetyl group of acetyl-TPP is partitioned by transfer to both CoASH and water. When the E{sub 1} component is incubated with ({sup 14}C)fluoropyruvate, TPP, and dihydrolipoamide, ({sup 14}C)acetyldihydrolipoamide is produced. The formation of ({sup 14}C)acetyldihydrolipoamide was examined as a function of dihydrolipoamide concentration. A plot of the extent of acetyl group-partitioning to dihydrolipoamide as a function of 1/(dihydrolipoamide) shows 95 {plus minus} 2% acetyl group transfer to dihydrolipoamide when dihydrolipoamide concentration is extrapolated to infinity. It is concluded that acetyl-TPP is chemically competent as an intermediate for the pyruvate dehydrogenase complex catalyzed oxidative decarboxylation of pyruvate.
- Research Organization:
- Wisconsin Univ., Madison, WI (USA)
- OSTI ID:
- 5564220
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550201* -- Biochemistry-- Tracer Techniques
550501 -- Metabolism-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMIDES
BACTERIA
BIOCHEMICAL REACTION KINETICS
BIOSYNTHESIS
CARBON 14 COMPOUNDS
CARBOXYLIC ACIDS
COMPLEXES
ENZYMES
ESCHERICHIA COLI
ISOTOPE APPLICATIONS
KETO ACIDS
KINETICS
LABELLED COMPOUNDS
METABOLISM
MICROORGANISMS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
OXIDOREDUCTASES
OXYGEN COMPOUNDS
PHOSPHORUS COMPOUNDS
PYROPHOSPHATES
PYRUVIC ACID
REACTION KINETICS
REAGENTS
SUBSTRATES
SYNTHESIS
TRACER TECHNIQUES
550501 -- Metabolism-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMIDES
BACTERIA
BIOCHEMICAL REACTION KINETICS
BIOSYNTHESIS
CARBON 14 COMPOUNDS
CARBOXYLIC ACIDS
COMPLEXES
ENZYMES
ESCHERICHIA COLI
ISOTOPE APPLICATIONS
KETO ACIDS
KINETICS
LABELLED COMPOUNDS
METABOLISM
MICROORGANISMS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
OXIDOREDUCTASES
OXYGEN COMPOUNDS
PHOSPHORUS COMPOUNDS
PYROPHOSPHATES
PYRUVIC ACID
REACTION KINETICS
REAGENTS
SUBSTRATES
SYNTHESIS
TRACER TECHNIQUES