title: Crystallization and preliminary X-ray analysis of native and selenomethionine 2-hydroxybiphenyl 3-monooxygenase.
abstract: 2-hydroxybiphenyl 3-monooxygenase (HbpA; EC 1.14.13.44) from Pseudomonas azelaica HBP1 was produced in Escherichia coli both as native and SeMet-labelled protein. The two enzymes were purified to homogeneity and crystallized by the hanging-drop vapour-diffusion method. The crystals belong to the monoclinic space group C2, with unit-cell parameters a = 108.6, b = 196.8, c = 79.3 A, beta = 97.7 degrees for the native protein and a = 108.3, b = 196.8, c = 79.0 A, beta = 97.8 degrees for SeMet HbpA. Crystal-packing considerations led to the assumption of two HbpA subunits per asymmetric unit, which corresponds to a V(M) value of 3.3 A(3) Da(-1) and a solvent content of 62%. The crystals were radiation-sensitive and only had a lifespan of about 120 s when exposed to synchrotron radiation on an undulator beamline. To obtain complete data sets, data were collected from 23 native and 26 derivative crystals. The high-resolution limit was 2.0 A for native and 2.25 A for SeMet HbpA.
authors.affiliation: Institute of Biotechnology, Swiss Federal Institute of Technology, ETHZ, Zurich, Switzerland.
authors.forename: Andreas
authors.initials: A
authors.lastname: Meyer
authors.forename: Daniel
authors.initials: D
authors.lastname: Tanner
authors.forename: Andreas
authors.initials: A
authors.lastname: Schmid
authors.forename: David F
authors.initials: DF
authors.lastname: Sargent
authors.forename: Hans-Peter E
authors.initials: HP
authors.lastname: Kohler
authors.forename: Bernard
authors.initials: B
authors.lastname: Witholt
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000737:chemistry*; D011549:Pseudomonas / Q000737:chemistry*; D012645:Selenomethionine / Q000737:chemistry / Q000378:metabolism; D014961:X-Ray Diffraction
chemical_list: D012645:Selenomethionine; D006899:Mixed Function Oxygenases; C058145:2-hydroxybiphenyl 3-monooxygenase