Biophysical Characterization of the Nucleoside Diphosphate Kinase of Leishmania major and Effect of the P95S Mutation

Autor(es)

Vieira, Plinio S. ^{[1, 2]} ; de Jesus Santos, Ana P. ^[2] ; de Oliveira, Arthur H. C. ^[2]

Afiliação do(s) autor(es)

^[1] CNPEM, LNBio, Campinas, SP - Brazil

^[2] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto FF, Dept Quim, BR-14040901 Ribeirao Preto, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: PROTEIN AND PEPTIDE LETTERS; v. 23, n. 2, p. 99-106, 2016.
• Citações Web of Science: 0

Resumo

Nucleoside diphosphate kinases (NDK; EC 2.7.4.6) are enzymes required for maintaining intracellular levels of nucleosides triphosphates (NTP) through transfer the gamma- phosphoryl group from a NTP to a NDP. The enzyme is associated with several biological functions including prevention of host ATP-mediated cytolysis during pathogenic infections. Here we present the biophysical characterization of NDK from Leishmania major and the effect of a mutation on the protein structure in solution. The structural stability was analyzed since this secreted protein may act in different microenvironments at various stages of the parasite life cycle. LmNDK and P95S mutant were subjected to denaturation with pH and guanidine. Structural transitions were monitored by circular dichroism and intrinsic fluorescence tryptophan emission. Our results showed that the LmNDK is more structurally stable than other described NDKs and that the catalytically active P95S mutant in the Kpn loop presented a decrease in protein stability, indicating the importance of this proline for maintenance of the LmNDK structure. (AU)

• Processo FAPESP: 11/20569-2 - Papel de aminoácidos do sítio ativo e da interface oligomérica na estrutura e função da nucleosídeo difosfato kinase de Leishmania major
• Beneficiário: Arthur Henrique Cavalcante de Oliveira
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 07/06755-2 - Caracterização estrutural e funcional de proteínas envolvidas na virulência do protozoário parasita Leishmania major
• Beneficiário: Arthur Henrique Cavalcante de Oliveira
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 10/03761-4 - Caracterização cinética e efeito de mutações na interface oligomérica da Nucleosídeo Difosfato Kinase de Leishmania major.
• Beneficiário: Plínio Salmazo Vieira
• Modalidade de apoio: Bolsas no Brasil - Mestrado