Biochemical characterization of recombinant nucleoside hydrolase from Mycobacterium tuberculosis H37Rv.

摘要: Tuberculosis (TB) is a major global health threat. There need for the development of more efficient drugs sterilization disease’s causative agent, Mycobacterium tuberculosis (MTB). A comprehensive understanding bacilli’s nucleotide metabolic pathways could aid in new anti-mycobacterial drugs. Here we describe expression and purification recombinant iunH-encoded nucleoside hydrolase from MTB (MtIAGU-NH). Glutaraldehyde cross-linking results indicate that MtIAGU-NH predominates as monomer, presenting varied oligomeric states depending upon binding ligands. Steady-state kinetics show has broad substrate specificity, accepting inosine, adenosine, guanosine, uridine substrates. Inosine adenosine displayed positive homotropic cooperativity kinetics, whereas guanosine hyperbolic saturation curves. Measurements ribose to by fluorescence spectroscopy suggest two pre-existing forms enzyme prior ligand association. The intracellular concentrations uridine, hypoxanthine, uracil were determined thermodynamic parameters estimated. Thermodynamic activation (Ea, ΔG#, ΔS#, ΔH#) MtIAGU-NH-catalyzed chemical reaction are presented. Results mass spectrometry, isothermal titration calorimetry (ITC), pH-rate profile experiment, multiple sequence alignment, molecular docking experiments also These data should contribute our biological role played MtIAGU-NH.