title: Expression, purification and preliminary crystallographic analysis of the N-terminal domain of Trypanosoma brucei BILBO1.
abstract: Trypanosoma brucei is a unicellular parasite that causes sleeping sickness in sub-Saharan Africa. It has a unique flagellar pocket (FP) at the base of the single flagellum. The FP is the sole site for endocytosis and exocytosis activity and plays crucial roles in the defence of the cell against the host immune response. In the neck region of the FP is an electron-dense material termed the flagellar pocket collar (FPC). T. brucei BILBO1 (TbBILBO1) was the first cytoskeletal protein to be characterized in the FPC. This protein is highly conserved among trypanosomatids and is essential for FP biogenesis. Structural information is needed to better understand the molecular mechanism of TbBILBO1 function in the cell. Here, the expression, purification and preliminary crystallographic analysis of the N-terminal domain of TbBILBO1 are reported. The protein was overexpressed in Escherichia coli strain BL21 (DE3), purified by multi-step chromatography and crystallized using the vapour-diffusion method. The crystal diffracted to 1.69 Å resolution and belonged to space group P21, with unit-cell parameters a = 29.69, b = 50.80, c = 37.22 Å, β = 94.61°. There was one molecule in the asymmetric unit.
authors.affiliation: Max F. Perutz Laboratories, Medical University of Vienna, Dr Bohr-gasse 9, 1030 Vienna, Austria.
authors.forename: Keni
authors.initials: K
authors.lastname: Vidilaseris
authors.affiliation: Max F. Perutz Laboratories, Medical University of Vienna, Dr Bohr-gasse 9, 1030 Vienna, Austria.
authors.forename: Gang
authors.initials: G
authors.lastname: Dong
mesh_terms: D018360:Crystallography, X-Ray; D005786:Gene Expression Regulation; D017434:Protein Structure, Tertiary / Q000502:physiology; D015800:Protozoan Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification*; D014346:Trypanosoma brucei brucei
chemical_list: D015800:Protozoan Proteins
keywords: BILBO1; Trypanosoma brucei; flagellar pocket