Molecular weight and coenzyme content of pyruvate decarboxylase from brewer's yeast

Ullrich, Johannes; Wittorf, John H.; Gubler, C. J.

doi:10.1016/s0926-6593(66)80017-6

Cited by 57 publications

(11 citation statements)

References 11 publications

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“…I n the second TOW from the bottom (Fig.7) the helix is right-handed in a t least 3 turns from the right of the picture, and then shifts to the left-handed towards the left. (7) x 380000, (8) X 380000, (9) In some of the plate-shaped crystals, areas of less organized pattern occurred (Fig. 6 -8).…”

Section: Electron Microscopymentioning

confidence: 99%

The pH 6 Acetolactate‐Forming Enzyme from Aerobacter aerogenes

Størmer

Solberg²,

Hovig³

1969

European Journal of Biochemistry

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The diffusion and sedimentation constants of the pH 6 acetolactate‐forming enzyme, determined by analytical ultracentrifugation, were used to calculate a molecular weight of 200 000 for the enzyme. The amino acid composition of the enzyme has been worked out, and 3 molecules of cocarboxylase, and 6 atoms of phosphorus were found to be tightly bound to one molecule of the enzyme. In addition, the effect of divalent cations was studied, results indicated that the enzyme has a requirement for manganese. An absolute requirement for cocarboxylase and manganese was not established. Two crystalline forms of the enzyme were studied by electron microscopy using negative staining technique. The plate‐shaped crystals were composed of rows of particles measuring approximately 100 × 60 Å. These particles consisted of sub‐units, and in some areas a helical organization of the crystal could be seen. The molecular organization was different in the two different crystalline forms of the enzyme.

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Section: Electron Microscopymentioning

confidence: 99%

The pH 6 Acetolactate‐Forming Enzyme from Aerobacter aerogenes

Størmer

Solberg²,

Hovig³

1969

European Journal of Biochemistry

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“…Pyruvate decarboxylase is widespread in eukaryotes, including plants and yeast, but is relatively rare in prokaryotes, having been found only in Sarcina ventriculi (Stephenson & Dawes,197 l), Zymomonas mobilis (Hoppner & Doelle, 1983), Acetobacter peroxydans (De Ley & Schell, 1959), Acetobacter suboxydans (King & Cheldelin, 1954) and Erwinia amylovora (Haq & Dawes,197 1). Purification of pyruvate decarboxylase has been achieved from two micro-organisms, Saccharomyces cerevisiae (Ullrich et al, 1966) and 2. mobilis (Hoppner & Doelle, 1983;Neale et al, 1987a).…”

Section: Introductionmentioning

confidence: 99%

Purification and characterization of pyruvate decarboxylase from Sarcina ventriculi

Lowe

Zeikus²

1992

Journal of General Microbiology

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Pyruvate decarboxylase from the obligate anaerobe Sarcina ventricufi was purified eightfold. The subunit M, was 57000 3OOO as estimated from SDS-PAGE, and the native M, estimated by gel filtration on a Superose 6 column was 240000, indicating that the enzyme is a tetramer. The M, values are comparable to those for pyruvate decarboxylase from Zymomonas mobifis and Saccharomyces cereoisiue, which are also tetrameric enzymes. The enzyme was oxygen stable, and had a pH optimum within the range 6.3-6.7. It displayed sigmoidal kinetics for pyruvate, with a S,, of 13 mM, kinetic properties also found for pyruvate decarboxylase from yeast and differing from the Michaelis-Menten kinetics of the enzyme from 2 . mobilis. No activators were found. p-Chloromercuribenzoate inhibited activity and the inhibition was reversed by the addition of dithiothreitol, indicating that cysteine is important in the active site. The N-terminal amino acid sequence of pyruvate decarboxylase was more similar to the sequence of S. cerevisiue than 2 . mobifis pyruvate decarboxylase.

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“…The structure of yeast PDC was resolved to 2.3 Å resolution [4]. The enzyme was described as a tetramer with a molecular mass of 240 000 Da [5,6]. The tetrameric molecule is a dimer of dimers that binds four molecules ThDP and four magnesium ions [4].…”

mentioning

confidence: 99%

Active oligomeric states of pyruvate decarboxylase and their functional characterization

Killenberg-Jabs

Jabs²,

Lilie

et al. 2001

European Journal of Biochemistry

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Homomeric pyruvate decarboxylase (E.C 4.1.1.1) from yeast consists of dimers and tetramers under physiological conditions, a K(d) value of 8.1 microM was determined by analytical ultracentrifugation. Dimers and monomers of the enzyme could be populated by equilibrium denaturation using urea as denaturant at defined concentrations and monitored by a combination of optical (fluorescence and circular dichroism) and hydrodynamic methods (analytical ultracentrifugation). Dimers occur after treatment with 0.5 M urea, monomers with 2.0 M urea independent of the protein concentration. The structured monomers are catalytically inactive. At even higher denaturant concentrations (6 M urea) the monomers unfold. The contact sites of two monomers in forming a dimer as the smallest enzymatically active unit are mainly determined by aromatic amino acids. Their interactions have been quantified both by structure-theoretical calculations on the basis of the X-ray crystallography structure, and experimentally by binding of the fluorescent dye bis-ANS. The contact sites of two dimers in tetramer formation, however, are mainly determined by electrostatic interactions. Homomeric pyruvate decarboxylase (PDC) is activated by its substrate pyruvate. There was no difference in the steady-state activity (specific activity) between dimers and tetramers. The activation kinetics of the two oligomeric states, however, revealed differences in the dissociation constant of the regulatory substrate (K(a)) by one order of magnitude. The tetramer formation is related to structural consequences of the interaction transfer in the activation process causing an improved substrate utilization.

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Molecular weight and coenzyme content of pyruvate decarboxylase from brewer's yeast

Cited by 57 publications

References 11 publications

The pH 6 Acetolactate‐Forming Enzyme from Aerobacter aerogenes

The pH 6 Acetolactate‐Forming Enzyme from Aerobacter aerogenes

Purification and characterization of pyruvate decarboxylase from Sarcina ventriculi

Active oligomeric states of pyruvate decarboxylase and their functional characterization

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