Open AccessCrystallization and preliminary X‐ray diffraction analysis of selenophosphate synthetases from Trypanosoma brucei and Leishmania major
Author(s) -
Faim Lívia Maria,
e Silva Ivan Rosa,
Dias Marcio Vinicius Bertacine,
Pereira Humberto D'Muniz,
BrandaoNeto José,
da Silva Marco Túlio Alves,
Thiemann Otavio Henrique
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113014632
Subject(s) - trypanosoma brucei , crystallization , selenium , leishmania , biochemistry , chemistry , x ray crystallography , biology , diffraction , organic chemistry , gene , physics , parasite hosting , world wide web , computer science , optics
Selenophosphate synthetase (SPS) plays an indispensable role in selenium metabolism, being responsible for catalyzing the activation of selenide with adenosine 5′‐triphosphate (ATP) to generate selenophosphate, the essential selenium donor for selenocysteine synthesis. Recombinant full‐length Leishmania major SPS ( Lm SPS2) was recalcitrant to crystallization. Therefore, a limited proteolysis technique was used and a stable N‐terminal truncated construct (ΔN‐ Lm SPS2) yielded suitable crystals. The Trypanosoma brucei SPS orthologue ( Tb SPS2) was crystallized by the microbatch method using paraffin oil. X‐ray diffraction data were collected to resolutions of 1.9 Å for ΔN‐ Lm SPS2 and 3.4 Å for Tb SPS2.