title: Cloning, expression, purification, crystallization and X-ray crystallographic analysis of Rv3168 from Mycobacterium tuberculosis H37Rv.
abstract: Tuberculosis is a widespread and deadly infectious disease, with one third of the human population already being infected. Aminoglycoside antibiotics have become less effective in recent years owing to antibiotic resistance, which arises primarily through enzymatic modification of the antibiotics. The gene product Rv3168, a putative aminoglycoside phosphotransferase (APH), from Mycobacterium tuberculosis was crystallized using the sitting-drop vapour-diffusion method in the presence of 0.2 M calcium acetate, 0.1 M Tris-HCl pH 7.0 and 20% PEG 3000 at 295 K. X-ray diffraction data were collected to a maximum resolution of 1.67 Å on a synchrotron beamline. The crystal belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 56.74, b = 62.37, c = 103.61 Å. With one molecule per asymmetric unit, the crystal volume per unit protein weight (V(M)) is 2.91 Å(3) Da(-1). The structure was solved by the single-wavelength anomalous dispersion method and refinement of the selenomethionine structure is in progress.
authors.affiliation: Pohang Accelerator Laboratory, Pohang University of Science and Technology, Pohang, Kyungbuk 790-784, Republic of Korea.
authors.forename: Sangwoo
authors.initials: S
authors.lastname: Kim
authors.forename: Chi My Thi
authors.initials: CM
authors.lastname: Nguyen
authors.forename: Seung Joo
authors.initials: SJ
authors.lastname: Yeo
authors.forename: Jae Woo
authors.initials: JW
authors.lastname: Ahn
authors.forename: Eun Jung
authors.initials: EJ
authors.lastname: Kim
authors.forename: Kyung Jin
authors.initials: KJ
authors.lastname: Kim
mesh_terms: D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D015870:Gene Expression; D008958:Models, Molecular; D009169:Mycobacterium tuberculosis / Q000201:enzymology*; D010770:Phosphotransferases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D017434:Protein Structure, Tertiary
chemical_list: D010770:Phosphotransferases