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Functional analysis of drug resistance-associated mutations in the Trypanosoma brucei adenosine transporter 1 (TbAT1) and the proposal of a structural model for the protein.
Munday, Jane C; Tagoe, Daniel N A; Eze, Anthonius A; Krezdorn, Jessica A M; Rojas López, Karla E; Alkhaldi, Abdulsalam A M; McDonald, Fiona; Still, Jennifer; Alzahrani, Khalid J; Settimo, Luca; De Koning, Harry P.
Afiliação
  • Munday JC; Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, G12 8TA, UK.
  • Tagoe DN; Wellcome Trust Centre for Molecular Parasitology, University of Glasgow, Glasgow, UK.
  • Eze AA; Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, G12 8TA, UK.
  • Krezdorn JA; Department of Laboratory Technology, University of Cape Coast, Cape Coast, Ghana.
  • Rojas López KE; Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, G12 8TA, UK.
  • Alkhaldi AA; Department of Medical Biochemistry, College of Medicine, University of Nigeria, Enugu Campus, Enugu, Nigeria.
  • McDonald F; Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, G12 8TA, UK.
  • Still J; Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, G12 8TA, UK.
  • Alzahrani KJ; Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, G12 8TA, UK.
  • Settimo L; Department of Biology, College of Science, Aljouf University, Sakaka, Saudi Arabia.
  • De Koning HP; Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, G12 8TA, UK.
Mol Microbiol ; 96(4): 887-900, 2015 May.
Article em En | MEDLINE | ID: mdl-25708978
The Trypanosoma brucei aminopurine transporter P2/TbAT1 has long been implicated in the transport of, and resistance to, the diamidine and melaminophenyl arsenical classes of drugs that form the backbone of the pharmacopoeia against African trypanosomiasis. Genetic alterations including deletions and single nucleotide polymorphisms (SNPs) have been observed in numerous strains and clinical isolates. Here, we systematically investigate each reported mutation and assess their effects on transporter function after expression in a tbat1(-/-) T. brucei line. Out of a set of six reported SNPs from a reported 'resistance allele', none significantly impaired sensitivity to pentamidine, diminazene or melarsoprol, relative to the TbAT1-WT allele, although several combinations, and the deletion of the codon for residue F316, resulted in highly significant impairment. These combinations of SNPs, and ΔF316, also strongly impaired the uptake of [(3)H]-adenosine and [(3)H]-diminazene, identical to the tbat1(-/-) control. The TbAT1 protein model predicted that residues F19, D140 and F316 interact with the substrate of the transporter. Mutation of D140 to alanine resulted in an inactive transporter, whereas the mutation F19A produced a transporter with a slightly increased affinity for [(3)H]-diminazene but reduced the uptake rate. The results presented here validate earlier hypotheses of drug binding motifs for TbAT1.
Assuntos

Texto completo: 1 Coleções: 01-internacional Contexto em Saúde: 3_ND Base de dados: MEDLINE Assunto principal: Tripanossomicidas / Trypanosoma brucei brucei / Modelos Moleculares / Proteínas de Transporte de Nucleosídeos Tipo de estudo: Prognostic_studies / Risk_factors_studies Idioma: En Revista: Mol Microbiol Ano de publicação: 2015 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Contexto em Saúde: 3_ND Base de dados: MEDLINE Assunto principal: Tripanossomicidas / Trypanosoma brucei brucei / Modelos Moleculares / Proteínas de Transporte de Nucleosídeos Tipo de estudo: Prognostic_studies / Risk_factors_studies Idioma: En Revista: Mol Microbiol Ano de publicação: 2015 Tipo de documento: Article