Structure of the Mycobacterium tuberculosis cPknF and conformational changes induced in forkhead-associated regulatory domains

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Cabarca, Sindy ^{[1, 2, 3]} ; de Souza, Maximilia Frazao ^[4] ; de Oliveira, Andrew Albert ^[5] ; Vignoli Muniz, Gabriel S. ^[6] ; Lamy, M. Teresa ^[6] ; dos Reis, Caio Vinicius ^[7] ; Takarada, Jessica ^[7] ; Effer, Brian ^{[8, 9]} ; Souza, Lucas Santos ^[2] ; de la Torre, Lilia Iriarte ^{[1, 2, 3]} ; Counago, Rafael ^[7] ; Pinto Oliveira, Cristiano Luis ^[4] ; Balan, Andrea ^[2]

Afiliação do(s) autor(es)

^[1] Univ Estadual Campinas, Programa Posgrad Genet, BR-13083862 Campinas, SP - Brazil

^[2] Univ Sao Paulo, Inst Ciencias Biomed, Dept Microbiol, Lab Biol Estrutural Aplicada LBEA, BR-05508000 Sao Paulo, SP - Brazil

^[3] Univ Sucre, Grp Invest Biomed, Sincelejo 700001, Sucre - Colombia

^[4] Univ Sao Paulo, Inst Fis, Dept Fis Expt, Grp Fluidos Complexos, BR-05508090 Sao Paulo, SP - Brazil

^[5] Univ Sao Paulo, Fac Med Ribeirao Preto, BR-14049900 Ribeirao Preto, SP - Brazil

^[6] Univ Sao Paulo, Inst Fis, Dept Fis Geral, BR-05508090 Sao Paulo - Brazil

^[7] Univ Estadual Campinas, Struct Genom Consortium, BR-13083886 Campinas, SP - Brazil

^[8] Univ La Frontera, Ctr Excellence Traslat Med CEMT, Temuco 01145 - Chile

^[9] Univ La Frontera, Sci & Technol Bioresource Nucleus BIOREN, Temuco 01145 - Chile

• Tipo de documento: Artigo Científico
• Fonte: CURRENT RESEARCH IN STRUCTURAL BIOLOGY; v. 3, p. 165-178, 2021.
• Citações Web of Science: 0

Resumo

Mycobacterium tuberculosis (Mtb) has 11 Serine-Threonine Protein Kinases (STPK) that control numerous physiological processes, including cell growth, cell division, metabolic flow, and transcription. PknF is one of the 11 Mtb STPKs that has, among other substrates, two FHA domains (FHA-1 and FHA-2) of the ATP-Binding Cassette (ABC) transporter Rv1747. Phosphorylation in T152 and T210 located in a non-structured linker that connects Rv1747 FHA domains is considerate to be the regulatory mechanism of the transporter. In this work, we resolved the three-dimensional structure of the PknF catalytic domain (cPknF) in complex with the human kinase inhibitor IKK16. cPknF is conserved when compared to other STPKs but shows specific residues in the binding site where the inhibitor is positioned. In addition, using Small Angle X-Ray Scattering analysis we monitored the behavior of the wild type and three FHA-phosphomimetic mutants in solution, and measured the cPknF affinity for these domains. The kinase showed higher affinity for the non-phosphorylated wild type domain and preference for phosphorylation of T152 inducing the rapprochement of the domains and significant structural changes. The results shed some light on the process of regulating the transporter's activity by phosphorylation and arises important questions about evolution and importance of this mechanism for the bacillus. (AU)

• Processo FAPESP: 18/20162-9 - Investigando a Patogênese e a Resistência à Drogas em Microrganismos - Caracterização e Controle de Transportadores ABC
• Beneficiário: Andrea Balan Fernandes
• Modalidade de apoio: Auxílio à Pesquisa - Regular