Espectrometria de massas combinada a agentes de ligação cruzada para a caracterização estrutural em solução da enzima Enoil-ACP-redutase de Mycobacterium tuberculosis

Abstract

Tuberculosis (TB), a disease mainly caused by the bacillus Mycobacterium tuberculosis (MTB), is considered a global public health problem. Infections caused by mycobacteria are generally difficult to treat because of its natural resistance to most antibiotics. This resistance is largely attributed to the formation of the cell wall. The InhA is part of the mycobacterial FAS II system which regulates the elongation of the fatty acid chain, which will compose the cell wall of the bacillus (mycolic acids). As the FAS II system is not present in mammals, this enzyme has been described as an important macromolecular target aiming the development of drugs with selective toxicity. The development of analytical techniques for studying proteins in solution by mass spectrometry combined with cross-linking has permitted access information about the primary, tertiary, and quaternary structure of the proteins. In this work, the technique of cross-linking combined with mass spectrometry was implemented for the characterization of the Mycobacterium tuberculosis enoyl-ACP reductase (InhA) which catalyzes the final essential enzymatic step in fatty acid elongation in the FAS II pathway, converting 2-trans-enoyl-ACP to acyl-ACP via an NADH-dependent reaction.