Molecular characterization of the E2 conjugating enzyme LinfUbc13 in Leishmania infantum

Autor(es)

da Silva, Eduardo Vagner Rodrigues ; Torres, Caroline ; Ribeiro, Hariel Nemamiah Escolarique ; de Correia, Camila Rolemberg Santana Travaglini Berti ; de Castro, Taissa de Oliveira ; Mancin, Giovanna da Costa ; Venancio, Mayla Gabriela Zanchetta ; Baqui, Munira Muhammad Abdel ; Teixeira, Felipe Roberti ; Gomes, Marcelo Damario

• Tipo de documento: Artigo Científico
• Fonte: Archives of Biochemistry and Biophysics; v. 764, p. 9-pg., 2024-12-19.

Resumo

UBC13 is an orthologue of Homo sapiens ubiquitin-conjugation E2 enzymes described in Leishmania mexicana, a null mutant lacking this gene cannot be produced, suggesting essential functions in this parasite. Leishmania infantum is an etiological agent of visceral leishmaniasis, the most severe type of disease that is potentially fatal if untreated. The ubiquitination process has been targeted for leishmanicidal compounds, indicating its essential function in parasite homeostasis. Therefore, the molecular characterization of the ubiquitination process may provide a better understanding of the molecular and cellular basis of leishmaniasis. Here, we characterized the gene LINF_350017900 in Leishmania infantum, which was named LinfUBC13, an E2 orthologue of UBC13 in Leishmania mexicana and the UBE2D family in Homo sapiens, sharing 72-74 % identity with UBE2D1, UBE2D2, and UBE2D3. LinfUbc13 contains conserved catalytic residues, including Cys86 and the HPN motif, which are essential for ubiquitin-conjugating activity. Structural analysis revealed a high similarity between LinfUbc13 and human UBE2D proteins, with a root-mean-square deviation (RMSD) of 0.4 & Aring;, suggesting conserved functions. Recombinant LinfUbc13 was expressed and shown to accept ubiquitin from E1, forming a thioester intermediate. Functional assays demonstrated that LinfUbc13 transfers ubiquitin to p53 through human HDM2 E3 ligase, confirming its role in ubiquitination. Subcellular localization showed that LinfUbc13 was distributed throughout the parasite cytoplasm. These findings highlight the conserved nature of the ubiquitin-proteasome system between Leishmania infantum and Homo sapiens, showing that LinfUbc13 is an E2 enzyme that plays a crucial role in parasitic development. (AU)

• Processo FAPESP: 21/10971-0 - Caracterização bioquímica das E3 ubiquitina-ligases do tipo CRL (Cullin RING-ligases) em Leishmania infantum
• Beneficiário: Camila Rolemberg Santana Travaglini Berti de Correia
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto
• Processo FAPESP: 22/02933-3 - Papel do sistema ubiquitina proteassoma na proliferação, diferenciação e infectividade de Leishmania infantum
• Beneficiário: Felipe Roberti Teixeira
• Modalidade de apoio: Auxílio à Pesquisa - Projeto Inicial
• Processo FAPESP: 22/16270-6 - Caracterização in silico do complexo E3 ubiquitina-ligases do tipo CRL1 (Cullin RING-ligases) em Leishmania infantum
• Beneficiário: Caroline Torres
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica
• Processo FAPESP: 23/17920-7 - Caracterização da interação do Trypanosoma cruzi com a maquinaria nuclear da célula hospedeira investigando as vias de reparo e medida de dano no DNA
• Beneficiário: Munira Muhammad Abdel Baqui
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 24/01732-0 - Caracterização funcional de proteínas do Sistema Ubiquitina Proteassoma de L.infantum
• Beneficiário: Taissa de Oliveira de Castro
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica