Leishmania donovani phosphofructokinase - Gene characterization, biochemical properties and structure-modelling studies
(2002) European Journal of Biochemistry — Vol. 269, n° 16, p. 3978-3989 (2002)
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- The characterization of the gene encoding Leishmania donovani phosphofructokinase (PFK) and the biochemical properties of the expressed enzyme are reported. L. donovani has a single PFK gene copy per haploid genome that encodes a polypeptide with a deduced molecular mass of 53 988 and a pI of 9.26. The predicted amino acid sequence contains a C-terminal tripeptide that conforms to an established signal for glycosome targeting. L. donovanii PFK showed most sequence similarity to inorganic pyrophosphate (PP)dependent PFKs, despite being ATP-dependent. It thereby resembles PFKs from other Kinetoplastida such as Trypanosoma brucei, Trypanoplasma borreli (characterized in this study), and a PFK found in Entamoeba histolytica. It exhibited hyperbolic kinetics with respect to ATP whereas the binding of the other substrate, fructose 6-phosphate, showed slight positive cooperativity. PPi, even at high concentrations, did not have any effect. AMP acted as an activator of PFK, shifting its kinetics for fructose 6-phosphate from slightly sigmoid to hyperbolic, and increasing considerably the affinity for this substrate, whereas GDP did not have any effect. Modelling studies and site-directed mutagenesis were employed to shed light on the structural basis for the AMP effector specificity and on ATP/PPi specificity among PFKs.
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López, C., Chevalier, N., Hannaert, V., Rigden, D., Michels, PAM., & Ramirez, J. (2002). Leishmania donovani phosphofructokinase - Gene characterization, biochemical properties and structure-modelling studies. European Journal of Biochemistry, 269(16), 3978-3989. https://doi.org/10.1046/j.1432-1033.2002.03086.x (Original work published 2002)