RESUMO
Sleeping sickness is a fatal disease caused by the protozoan parasite Trypanosoma brucei (Tb). Inosine-5'-monophosphate dehydrogenase (IMPDH) has been proposed as a potential drug target, since it maintains the balance between guanylate deoxynucleotide and ribonucleotide levels that is pivotal for the parasite. Here we report the structure of TbIMPDH at room temperature utilizing free-electron laser radiation on crystals grown in living insect cells. The 2.80 Å resolution structure reveals the presence of ATP and GMP at the canonical sites of the Bateman domains, the latter in a so far unknown coordination mode. Consistent with previously reported IMPDH complexes harboring guanosine nucleotides at the second canonical site, TbIMPDH forms a compact oligomer structure, supporting a nucleotide-controlled conformational switch that allosterically modulates the catalytic activity. The oligomeric TbIMPDH structure we present here reveals the potential of in cellulo crystallization to identify genuine allosteric co-factors from a natural reservoir of specific compounds.
Assuntos
Coenzimas/química , Cristalização , IMP Desidrogenase/química , Trypanosoma brucei brucei/enzimologia , Sequência de Aminoácidos , Animais , Sítios de Ligação , Domínio Catalítico , Clonagem Molecular , Guanosina Monofosfato , Modelos Moleculares , Conformação Proteica , Células Sf9 , Trypanosoma brucei brucei/genéticaRESUMO
The aim of this study was to determine the incidence of 18 environmental variables in the spatial distribution of 30 chorotypes (species groups with significantly similar distribution patterns) of dorylaimid and mononchid nematodes by means of logistic regression in a natural area in the southeastern Iberian Peninsula. Six variables (elevation, color chroma, clay content, nitrogen content, CaCO, and plant community associated) were the most important environmental factors that helped explain the distribution of chorotypes. The distribution of most chorotypes was characterized by some (one to three) environmental variables; only two chorotypes were characterized by five or more variables, and four have not been characterized.