Kinetic and Regulatory Properties of Heart Muscle Pyruvate Dehydrogenase

Wieland, O.; Jagow-Westermann, Bärbel; Stukowski, Barbara

doi:10.1515/bchm2.1969.350.1.329

Cited by 103 publications

(10 citation statements)

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“…Thus, phosphatase activity was determined by measuring the rate of reactivation of a constant amount of phospho pyruvate dehydrogenase : a mixture containing in a final volume of 20p.1, 50mU phospho pyruvate dehydrogenase, 0.2 pmol MgC1, and 5 p1 of phosphatase fraction was incubated in a sealed microcup (Netheler & Hinz, Hamburg-Eppendorf, Germany) for 5 min a t 25 "C. Thereafter a 10-p1 aliquot was assayed for pyruvate dehydrogenase activity spectrophotometrically with pyruvate as substrate as described earlier [9]. For measuring phosphatase activity in the crude heart muscle extracts the dismutation assay of Korkes et al [lo] standardized with purified pyruvate dehydrogenase was applied.…”

Section: Standard Phosphatase Assaymentioning

confidence: 99%

Purification and Characterization of Pyruvate‐Dehydrogenase Phosphatase from Pig‐Heart Muscle

Siess

Wieland

1972

European Journal of Biochemistry

109

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1. Pyruvate-dehydrogenase phosphatase was extensively purified from pig heart muscle. 2. The molecular weight was determined to be in the range between 92000 and 95000. By dodecylsulfate-gel electrophoresis there was no indication for a subunit structure of the protein.3. The phosphatase is dependent on Mg2f or Mn2+. Half-maximal activity was obtained, in histidine buffer, a t 2.5 mM MgC1, and 1.8 mM MnCI,, respectively.4. Fluoride a t a concentration of 0.6mM inhibited phosphatase activity by 50°/, in the presence of Mg2+, but not of Mn2+. The enzyme was not affected by alkylating thiol-reagents.5. Studies with sequestering agents suggest that pyruvate-dehydrogenase phosphatasc contains a metal which, besides Mg2+ or Mn2+, is essential for its function.The purified enzyme complex of mammalian pyruvate dehydrogenase is converted from the catalytically active form to the inactive form by ATPdependent phosphorylation [l --51. The enzyme responsible for the phosphate incorporation was called "pyruvate dehydrogenase kinase". Reactivation of the phospho pyruvate-dehydrogenase complex is catalyzed by a second enzyme entity, which was designated pyruvate-dehydrogenase phosphatase, since the pyruvate-dehydrogenase complex is concomitantly dephosphorylated during reactivation [ 1 , 3 -51.

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Section: Standard Phosphatase Assaymentioning

confidence: 99%

Purification and Characterization of Pyruvate‐Dehydrogenase Phosphatase from Pig‐Heart Muscle

Siess

Wieland

1972

European Journal of Biochemistry

109

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“…In preparations from both the patient and controls, the absence of added coenzyme was associated with 20-30% lower activity, whereas the addition of twice the usual amount of coenzyme caused no significant change (Table VI). Complete dissocation of pyruvate decarboxylase apoenzyme from the coenzyme has been shown usually to require alkaline conditions (27,28). Michaelis-Menton constants (Km) were derived from the curves shown in Fig.…”

Section: Metabolic Studiesmentioning

confidence: 99%

A defect in pyruvate decarboxylase in a child with an intermittent movement disorder

Blass¹,

Avigan²,

Uhlendorf³

1970

J. Clin. Invest.

247

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“…In 48 h starved or diabetic rats phosphorylation and inactivation of the complex inhibits pyruvate and glucose oxidation [2,3]. This is apparently effected by increased activity of PDH kinase [4-61.…”

Section: Introductionmentioning

confidence: 99%

The roles of intrinsic kinase and of kinase/activator protein in the enhanced phosphorylation of pyruvate dehydrogenase complex in starvation

1984

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Extracts of heart mitochondria from fed and from 48 h starved rats subjected to gel filtration on Sephacryl S-300 gave 4 major protein peaks. Pyruvate dehydrogenase complex eluted in the void volume and was assayed for intrinsic pyruvate dehydrogenase kinase activity which was increased approximately 3-fold by 48 h starvation of the rat. A second fraction, containing peaks 2 and 3 which overlapped, enhanced the activity of the intrinsic kinase and corresponds to ~n~/activator protein described previously. Its activity was increased X.5fold by starvation.

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Kinetic and Regulatory Properties of Heart Muscle Pyruvate Dehydrogenase

Cited by 103 publications

References 0 publications

Purification and Characterization of Pyruvate‐Dehydrogenase Phosphatase from Pig‐Heart Muscle

Purification and Characterization of Pyruvate‐Dehydrogenase Phosphatase from Pig‐Heart Muscle

A defect in pyruvate decarboxylase in a child with an intermittent movement disorder

The roles of intrinsic kinase and of kinase/activator protein in the enhanced phosphorylation of pyruvate dehydrogenase complex in starvation

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