title: Purification, crystallization and preliminary X-ray analysis of the periplasmic haem-binding protein HutB from Vibrio cholerae.
abstract: The mechanism of haem transport across the inner membrane of pathogenic bacteria is currently insufficiently understood at the molecular level and no information is available for this process in Vibrio cholerae. To obtain structural insights into the periplasmic haem-binding protein HutB from V. cholerae (VcHutB), which is involved in haem transport through the HutBCD haem-transport system, at the atomic level, VcHutB was cloned, overexpressed and crystallized using 1.6 M ammonium sulfate as a precipitant at pH 7.0. X-ray diffraction data were collected to 2.4 Å resolution on the RRCAT PX-BL-21 beamline at the Indus-2 synchrotron, Indore, India. The crystals belonged to space group P4₃2₁2, with unit-cell parameters a = b = 62.88, c = 135.8 Å. Matthews coefficient calculations indicated the presence of one monomer in the asymmetric unit, with an approximate solvent content of 45.02%. Molecular-replacement calculations with Phaser confirmed the presence of a monomer in the asymmetric unit.
authors: Department of Biotechnology, St Xavier's College, 30 Park Street, Kolkata 700 016, India.
authors: Shubhangi
authors: S
authors: Agarwal
authors: Department of Biotechnology, St Xavier's College, 30 Park Street, Kolkata 700 016, India.
authors: Maitree
authors: M
authors: Biswas
authors: Department of Biotechnology, St Xavier's College, 30 Park Street, Kolkata 700 016, India.
authors: Jhimli
authors: J
authors: Dasgupta
mesh_terms: D002352:Carrier Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D003460:Crystallization; D000081062:Heme-Binding Proteins; D006420:Hemeproteins / Q000737:chemistry* / Q000302:isolation & purification*; D019897:Periplasm / Q000737:chemistry*; D014734:Vibrio cholerae; D014961:X-Ray Diffraction
chemical_list: D002352:Carrier Proteins; D000081062:Heme-Binding Proteins; D006420:Hemeproteins
keywords: ABC transporter; Vibrio cholerae; iron uptake; periplasmic haem-binding protein