A biochemical and genetic study of Leishmania donovani pyruvate kinase

Gene
Will SandovalJose Luis Ramirez

Abstract

Here we present a biochemical and molecular biology study of the enzyme pyruvate kinase (PYK) from the parasitic protozoa Leishmania donovani. The PYK gene was cloned, mutagenised and over expressed and its kinetic parameters determined. Like in other kinetoplastids, L. donovani PYK is allosterically stimulated by the effector fructose 2,6 biphosphate and not by fructose 1,6 biphosphate. When the putative effector binding site of L. donovani PYK was mutagenised, we obtained two mutants with extreme kinetic behavior: Lys453Leu, which retained a sigmoidal kinetics and was little affected by the effector; and His480Gln, which deployed a hyperbolic kinetics that was not changed by the addition of the effector. Molecular Dynamics (MD) studies revealed that the mutations not only altered the effector binding site of L. donovani PYK but also changed the folding of its domain C.

References

Nov 24, 1979·Nucleic Acids Research·H C Birnboim, J Doly
Jan 25, 1992·Nucleic Acids Research·I Mikaelian, A Sergeant
Dec 5, 1993·Journal of Molecular Biology·A Sali, T L Blundell
Jan 1, 1993·Archives of Biochemistry and Biophysics·A Ponte-SucreJ L Ramirez
Jan 19, 2006·Clinical Microbiology Reviews·Simon L CroftAlan H Fairlamb
Oct 17, 2006·Parasitology Today·A G Tielens, J J Van Hellemond
Jun 19, 2007·Nature Genetics·Christopher S PeacockMatthew Berriman

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Citations

Jan 5, 2011·Antimicrobial Agents and Chemotherapy·José Ignacio ManzanoFrancisco Gamarro

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