Production, purification, crystallization and preliminary X-ray diffraction studies of the nucleoside diphosphate kinase b from Leishmania major.
Nucleoside diphosphate kinases (NDKs; EC 2.7.4.6) play an essential role in the synthesis of nucleotides from intermediates in the salvage pathway in all parasitic trypanosomatids and their structural studies will be instrumental in shedding light on the biochemical machinery involved in the parasite life cycle and host-parasite interactions. In this work, NDKb from Leishmania major was overexpressed in Escherichia coli, purified to homogeneity and crystallized using the sitting-drop vapour-diffusion method. The NDK crystal diffracted to 2.2 angstrom resolution and belonged to the trigonal crystal system, with unit-cell parameters a = 114.2, c = 93.9 angstrom. Translation-function calculations yielded an unambiguous solution in the enantiomorphic space group P3(2)21.
Tonoli
Celisa Caldana Costa
CC
Center for Structural Molecular Biology, Brazilian Association for Synchrotron Light Technology, Campinas-SP, Brazil.
Vieira
Plinio Salmazo
PS
Ward
Richard John
RJ
Arni
Raghuvir Krishnaswamy
RK
de Oliveira
Arthur Henrique Cavalcante
AH
Murakami
Mario Tyago
MT
D000595:Amino Acid Sequence; D000818:Animals; D003460:Crystallization; D018360:Crystallography, X-Ray; D018320:Leishmania major / Q000201:enzymology*; D008969:Molecular Sequence Data; D054778:NM23 Nucleoside Diphosphate Kinases / Q000737:chemistry* / Q000235:genetics; D015800:Protozoan Proteins / Q000737:chemistry* / Q000235:genetics; D014961:X-Ray Diffraction
D054778:NM23 Nucleoside Diphosphate Kinases; D015800:Protozoan Proteins