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Increased Stability and Catalytic Efficiency of Yeast Hexokinase Upon Interaction with Zwitterionic Micelles. Kinetics and Conformational Studies

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Abstract

The effect of ligands (glucose, ATP and Mg2+) and zwitterionic micellesof lysophosphatidylcholine (LPC) or N-hexadecyl-N,N-dimethyl-3-ammoniumpropanesulfonate (HPS) in the yeast hexokinase (HK) stability was studied at35°C. The thermal inactivation kinetics followed one-exponentialdecay. The effect of ligands on protecting the enzyme against inactivationfollowed the order: glucose>glucose/Mg2+>ATP/Mg2+≌Mg2+≌bufferonly. Both LPC and HPS micelles increased the enzyme stability only whenthe incubation medium contained glucose or glucose/Mg2+,suggesting that the protein conformation is a key prerequisite for theenzyme-micelle interaction to take place. This enzyme-micelle interactionresulted in an increased catalytic efficiency (with a decrease in Km forATP and increase in Vmax as well as in changes on the tertiary (intrinsicfluorescence) structure of the yeast hexokinase.

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Authors and Affiliations

  1. Departamento de Bioquímica Médica, ICB/CCS, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, CEP 21941-590, Brazil

    Rodrigo Guerra

  2. Departamento de Bioquímica Médica, ICB/UFRJ, Prédio do CCS, bloco E, sala 38, Ilha do Fundão, Rio de Janeiro, RJ, CEP 21941-590, Brazil

    M. Lucia Bianconi

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  1. Rodrigo Guerra
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  2. M. Lucia Bianconi
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Guerra, R., Bianconi, M.L. Increased Stability and Catalytic Efficiency of Yeast Hexokinase Upon Interaction with Zwitterionic Micelles. Kinetics and Conformational Studies. Biosci Rep 20, 41–49 (2000). https://doi.org/10.1023/A:1005583117296

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