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The putative effector-binding site of Leishmania mexicana pyruvate kinase studied by site-directed mutagenesis.

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Hannaert, Véronique [UCL] Yernaux, Cédric [UCL] Rigden, Daniel J Fothergill-Gilmore, Linda A. Opperdoes, Frederik [UCL] Michels, Paulus [UCL]

The activity of pyruvate kinase of Leishmania mexicana is allosterically regulated by fructose 2,6-bisphosphate (F-2,6-P(2)), contrary to the pyruvate kinases from other eukaryotes that are usually stimulated by fructose 1,6-bisphosphate (F-1,6-P(2)). Based on the comparison of the three-dimensional structure of Saccharomyces cerevisiae pyruvate kinase crystallized with F-1,6-P(2) present at the effector site (R-state) and the L. mexicana enzyme crystallized in the T-state, two residues (Lys453 and His480) were proposed to bind the 2-phospho group of the effector. This hypothesis was tested by site-directed mutagenesis. The allosteric activation by F-2,6-P(2) appeared to be entirely abrogated in the mutated enzymes confirming our predictions.

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Bibliographic reference Hannaert, Véronique ; Yernaux, Cédric ; Rigden, Daniel J ; Fothergill-Gilmore, Linda A. ; Opperdoes, Frederik ; et. al. The putative effector-binding site of Leishmania mexicana pyruvate kinase studied by site-directed mutagenesis.. In: FEBS letters, Vol. 514, no. 2-3, p. 255-9 (2002)
Permanent URL http://hdl.handle.net/2078.1/9128