Autor(es)

Rosa, Luis Victor Santana ; Blaszczyk, Beata ; Blundell, Tom ; Dias, Marcio Vinicius Bertacine

• Tipo de documento: Artigo Científico
• Fonte: International Journal of Biological Macromolecules; v. 330, p. 9-pg., 2025-09-30.

Resumo

Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis (Mtb) and is one of the leading causes of death worldwide. This disease is typically treated by combining several antimicrobials for extended periods, which can lead to treatment interruptions by patients and promote the emergence of multidrug-resistant strains, necessitating the use of alternative or second-line drugs. In this perspective, dihydrodipicolinate synthase (DapA) from M. tuberculosis (MtDapA), which catalyzes the aldol condensation between pyruvate and aspartatesemialdehyde (ASA) to produce dihydrodipicolinate, is an essential enzyme in Mtb for the production of Llysine and meso-diaminopimelate (mDAP). Through crystallographic assays, we have determined the structure of MtDapA in complex with its substrate, pyruvate, covalently bonded through a Schiff base to the catalytic L-lysine at a resolution of 1.5 & Aring;. Through structural analysis, we describe the arrangement of interactions between the active site amino acid residues and pyruvate, providing insight into the binding mode of this molecule. In addition, we performed further biophysical assays, including differential scanning fluorimetry (DSF) and isothermal titration calorimetry (ITC), to obtain insights into the pyruvate affinity and the potential role of Llysine and mDAP as allosteric regulators of MtDapA. However, in contrast to those observed in other orthologous enzymes, particularly those from Gram-negative bacteria, MtDapA does not have an affinity for L-lysine or mDAP. Consequently, this enzyme is not allosterically regulated by the products of this pathway. The results shown here provide evidence regarding the functioning of the enzyme regulatory mechanism and valuable structural features to aid in the future development of MtDapA inhibitors, which may be further explored in drug discovery campaigns against tuberculosis. (AU)

• Processo FAPESP: 21/10577-0 - Centro de Pesquisa em Biologia de Bactérias e Bacteriófagos (CEPID B3)
• Beneficiário: Shaker Chuck Farah
• Modalidade de apoio: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs
• Processo FAPESP: 22/12234-5 - Resistência a antimicrobianos em Mycobacterium tuberculosis: análise estrutural, biofísica e bioquímica de mutações missense
• Beneficiário: Marcio Vinicius Bertacine Dias
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 22/03967-9 - Caracterização funcional e estrutural preliminar da interação do domínio Transpeptidase PonA1 de Micobactérias com antibióticos beta-lactâmicos
• Beneficiário: Luís Victor Santana Rosa
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica