IBR   13079
INSTITUTO DE BIOLOGIA MOLECULAR Y CELULAR DE ROSARIO
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
DEVELOPMENT OF A HIGH THROUGHPUT ASSAY FOR THE SCREENING OF ACYL-COA CARBOXYLASE INHIBITORS
Autor/es:
BAZET LYONET BERNARDO; DIACOVICH LAUTARO; GAGO GABRIELA; GRAMAJO HUGO
Lugar:
Mar del Plata
Reunión:
Congreso; Reunión Anual Nº 51 (LI) de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2015
Institución organizadora:
Sociedad Argentina de Investigación en Bioquímica y Biología Molecular (SAIB)
Resumen:
Acyl-CoA carboxylases (ACCases) commit acyl-CoAs to the biosynthesis of lipids in Mycobacterium tuberculosis. This pathogen has several genes coding for ACCase subunits in its genome: three α subunits (accA1?3), six β subunits (accD1?6) and one ε subunit (accE5). ACCase 5 complex is formed by the biotinylated α subunit AccA3, the carboxyltransferase β subunit AccD5 and the small ε subunit AccE5. To gain insight about the metabolic relevance of this enzyme in mycobacteria, we obtained M. smegmatis mutants in accD5-accE5, the two subunits specifically associated with ACCase 5. The analysis of this conditional mutant demonstrated that AccD5 and AccE5 are part of an essential ACCase involved in lipid biosynthesis, and proposed ACCase 5 as an attractive target for tuberculosis drug discovery. In this sense, we developed an enzyme-based assay to identify inhibitors of ACCase 5, and we optimized it for high throughput screening. We used this assay towards a library containing 11,000 compounds and found 34 candidates. We further analyzed these candidates by conventional methods and we found 6 compounds that inhibit ACCase 5 with different potency and now need to be characterized in more detail. These results validated the high throughput screening assay as a powerful tool for identifying novel enzyme inhibitors that could be developed as anti-tuberculosis drugs.