Expression, purification and preliminary crystallographic analysis of oligopeptidase B from Trypanosoma brucei.
African sleeping sickness, also called trypanosomiasis, is a significant cause of morbidity and mortality in sub-Saharan Africa. Peptidases from Trypanosoma brucei, the causative agent, include the serine peptidase oligopeptidase B, a documented virulence factor and therapeutic target. Determination of the three-dimensional structure of oligopeptidase B is desirable to facilitate the development of novel inhibitors. Oligopeptidase B was overexpressed in Escherichia coli as an N-terminally hexahistidine-tagged fusion protein, purified using metal-affinity chromatography and crystallized using the hanging-drop vapour-diffusion technique in 7%(w/v) polyethylene glycol 6000, 1 M LiCl, 0.1 M bis-tris propane pH 7.5. Diffraction data to 2.7 angstroms resolution were collected using synchrotron radiation. The crystals belong to space group P3(1)21 or P3(2)21, with unit-cell parameters a = b = 124.5, c = 249.9 angstroms. A complete data set to 2.7 angstroms was collected using synchrotron radiation.
Rea
Dean
D
Department of Biological Sciences, University of Warwick, Gibbet Hill Road, Coventry CV4 7AL, England.
Hazell
Carole
C
Andrews
Norma W
NW
Morty
Rory E
RE
Fülöp
Vilmos
V
D000595:Amino Acid Sequence; D000818:Animals; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D011487:Protein Conformation; D015800:Protozoan Proteins / Q000737:chemistry / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D014346:Trypanosoma brucei brucei / Q000201:enzymology*
D010446:Peptide Fragments; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D012697:Serine Endopeptidases; C090703:oligopeptidase B