Skeletal-muscle UDP-glucose pyrophosphorylase is inactivated by reaction with 2-ethoxy-N-(ethoxy-carbonyl)-1,2-dihydroquinoline (EEDQ) and 1-(3-dimethylaminopropyl-3-ethylcarbodi-imide (EDAC), two reagents specific for carboxylate groups. The former reagent is a more effective inactivator than EDAC. Although no evidence of reversible enzyme-reagent complexes of the affinity-labelling type was obtained by kinetic analysis of the inactivation, the selective protection of UDP-glucose pyrophosphorylase activity against inactivation by EEDQ in the presence of uridine substrates is indicative of an active-site-directed effect. The results are consistent with the hypothesis that EEDQ modifies a single carboxylate group located in a hydrophobic domain close to the substrate-binding site, leading to enzyme inactivation. In contrast, the reaction between UDP-glucose pyrophosphorylase and EDAC appears to involve a different region of the enzyme.
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December 1989
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Research Article|
December 15 1989
Inactivation of skeletal-muscle UDP-glucose pyrophosphorylase by reaction with carboxylate-directed reagents
M Signorini;
M Signorini
1Istituto di Chimica Biologica, Universitá di Ferrara, Via Borsari 46, 44100 Ferrara, Italy.
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C Ferrari;
C Ferrari
1Istituto di Chimica Biologica, Universitá di Ferrara, Via Borsari 46, 44100 Ferrara, Italy.
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E Mariotti;
E Mariotti
1Istituto di Chimica Biologica, Universitá di Ferrara, Via Borsari 46, 44100 Ferrara, Italy.
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F Dallocchio;
F Dallocchio
1Istituto di Chimica Biologica, Universitá di Ferrara, Via Borsari 46, 44100 Ferrara, Italy.
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C M Bergamini
C M Bergamini
1Istituto di Chimica Biologica, Universitá di Ferrara, Via Borsari 46, 44100 Ferrara, Italy.
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Publisher: Portland Press Ltd
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© 1989 London: The Biochemical Society
1989
Biochem J (1989) 264 (3): 799–804.
Citation
M Signorini, C Ferrari, E Mariotti, F Dallocchio, C M Bergamini; Inactivation of skeletal-muscle UDP-glucose pyrophosphorylase by reaction with carboxylate-directed reagents. Biochem J 15 December 1989; 264 (3): 799–804. doi: https://doi.org/10.1042/bj2640799
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