Mycobacterium tuberculosis CysA2 is a dual sulfurtransferase with activity against thiosulfate and 3-mercaptopyruvate and interacts with mammalian cells

Autor(es)

Meza, A. N. ^{[1, 2]} ; Cambui, C. C. N. ^[1] ; Moreno, A. C. R. ^[3] ; Fessel, M. R. ^[1] ; Balan, A. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, LBEA, Inst Biomed Sci, Dept Microbiol, Appl Struct Biol Lab, Sao Paulo, SP - Brazil

^[2] Univ Estadual Campinas, UNICAMP, Postgrad Program Genet & Mol Biol, Inst Biol, Campinas, SP - Brazil

^[3] Univ Sao Paulo, Biomed Sci Inst, Dept Microbiol, Vaccine Dev Lab, Sao Paulo, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: SCIENTIFIC REPORTS; v. 9, NOV 14 2019.
• Citações Web of Science: 0

Resumo

Cyanide is a toxic compound that is converted to the non-toxic thiocyanate by a rhodanese enzyme. Rhodaneses belong to the family of transferases (sulfurtransferases), which are largely studied. The sulfur donor defines the subfamily of these enzymes as thiosulfate:cyanide sulfurtransferases or rhodaneses (TSTs) or 3-mercaptopyruvate sulfurtransfeases (MSTs). In Mycobacterium tuberculosis, the causative agent of tuberculosis, the gene Rv0815c encodes the protein CysA2, a putative uncharacterized thiosulfate:cyanide sulfurtransferase that belongs to the essential sulfur assimilation pathway in the bacillus and is secreted during infection. In this work, we characterized the functional and structural properties of CysA2 and its kinetic parameters. The recombinant CysA2 is a alpha/beta protein with two rhodanese-like domains that maintains the functional motifs and a catalytic cysteine. Sulfurtransferase activity was determined using thiosulfate and 3-mercaptopyruvate as sulfur donors. The assays showed K-m values of 2.89 mM and 7.02 mM for thiosulfate and 3-mercaptopyruvate, respectively, indicating the protein has dual activity as TST and MST. Immunological assays revealed that CysA2 interacted with pulmonary cells, and it was capable to activate macrophages and dendritic cells, indicating the stimulation of the immune response, which is important for its use as an antigen for vaccine development and immunodiagnostic. (AU)

• Processo FAPESP: 14/20921-6 - Identificação e desenvolvimento de novas moléculas contra Mycobacterium tuberculosis baseado na inibição de proteínas do metabolismo de enxofre utilizando a metodologia baseada em fragmentos
• Beneficiário: Andreia Navarro Cerone
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 15/14514-1 - Estudos Funcionais e Estruturais de Transportadores do Tipo ABC de Mycobacterium tuberculosis e Xanthomonas citri
• Beneficiário: Andrea Balan Fernandes
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 16/03196-1 - Expressão, purificação e caracterização cinética e antigênica da proteína CysA2, suposta enzima com atividade rhodanese-like envolvida no metabolismo de enxofre em Mycobacterium tuberculosis
• Beneficiário: Caio Cesar Nogueira Cambui
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica
• 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