Structural and kinetic characterization of Trypanosoma congolense pyruvate kinase

• Published in: Molecular and biochemical parasitology Vol. 236; p. 111263
• Main Authors: Pinto Torres, Joar Esteban, Yuan, Meng, Goossens, Julie, Versées, Wim, Caljon, Guy, Michels, Paul A., Walkinshaw, Malcolm D., Magez, Stefan, Sterckx, Yann G.-J.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.03.2020
• Subjects: Animals; Cattle; Cattle Diseases - parasitology; Enzyme kinetics; Glycolysis; Humans; Kinetics; Models, Structural; Protozoan Proteins - chemistry; Protozoan Proteins - metabolism; Pyruvate kinase; Pyruvate Kinase - chemistry; Pyruvate Kinase - metabolism; Trypanosoma congolense - metabolism; Trypanosomes; Trypanosomiasis, African - parasitology; X-ray crystallography; X-ray crystallography; Trypanosomes; Glycolysis; Pyruvate kinase; Enzyme kinetics
• ISSN: 0166-6851; 1872-9428
• DOI: 10.1016/j.molbiopara.2020.111263

•Effectors F16BP and F26BP increase the specificity constant of TcoPYK towards PEP.•Effector and substrate binding increase TcoPYK's thermal stability.•TcoPYK adopts an R-state conformation when bound by F16BP.•Citrate weakly inhibits the enzymatic activity of TcoPYK. Trypanosoma are blood-borne parasites and are the causative agents of neglected tropical diseases (NTDs) affecting both humans and animals. These parasites mainly rely on glycolysis for their energy production within the mammalian host, which is why trypanosomal glycolytic enzymes have been pursued as interesting targets for the development of trypanocidal drugs. The structure–function relationships of pyruvate kinases (PYKs) from trypanosomatids (Trypanosoma and Leishmania) have been well-studied within this context. In this paper, we describe the structural and enzymatic characterization of PYK from T. congolense (TcoPYK), the main causative agent of Animal African Trypanosomosis (AAT), by employing a combination of enzymatic assays, thermal unfolding studies and X-ray crystallography.