Pyruvatekinase from the hepatopancreas of the common shore crab, Carcinus maenas, was purified to a specific activity of 240 units/mg of protein in the assay conditions described. 2. In one method of purification the enzymic activity could be resolved into two fractions after chromatography on DEAE-cellulose. Fructose 1, 6-diphosphate was able to effect the conversion of one form (peak 1) into the second (peak 2). 3. In the presence of a saturating concentration of fructose 1, 6-diphosphate both forms of the enzyme were kinetically similar. 4. Polyacrylamide-gel electrophoresis of the enzyme 1 day after preparation showed a single protein band. On storage at least three protein bands became visible, all of which were associated with pyruvate kinase activity. 5. Chromatography of the enzyme on Sephadex G-200 indicated a mol.wt. of 247000, but in the presence of fructose 1, 6-diphosphate the elution volume of the enzyme increased corresponding to a mol.wt. of 193000. 6 Dissociation of the enzyme in sodium dodecyl sulphate and 2-mercaptoethanol followed by polyacrylamide-gel electrophoresis produced one major protein band with a mol.wt. of 55000.
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Research Article|
January 01 1976
Purification and properties of pyruvate kinase from the hepatopancreas of Carcinus maenas
I G Giles;
I G Giles
1Department of Physiology and Biochemistry, University of Southampton, Southampton S09 5NH, U.K.
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P C Poat;
P C Poat
1Department of Physiology and Biochemistry, University of Southampton, Southampton S09 5NH, U.K.
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K A Munday
K A Munday
1Department of Physiology and Biochemistry, University of Southampton, Southampton S09 5NH, U.K.
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Publisher: Portland Press Ltd
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© 1976 London: The Biochemical Society
1976
Biochem J (1976) 153 (1): 127–134.
Citation
I G Giles, P C Poat, K A Munday; Purification and properties of pyruvate kinase from the hepatopancreas of Carcinus maenas. Biochem J 1 January 1976; 153 (1): 127–134. doi: https://doi.org/10.1042/bj1530127
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