title: Three-dimensional model of Escherichia coli gyrase B subunit crystallized in two-dimensions on novobiocin-linked phospholipid films.
abstract: Two-dimensional crystals of the Escherichia coli DNA gyrase B subunit were obtained upon specific interactions with novobiocin linked phospholipid films. A three-dimensional surface model of the protein was generated by analysing images of tilted negatively stained crystals. The structure showed, at 2.5 to 3.0 nm resolution, two elongated arms organised as a V-shaped protein: the bottom of the V contains the novobiocin binding site, and the extremities of the arms mediate protein-protein interactions between the two monomers in the unit cell. Image analysis of frozen hydrated two-dimensional crystals resulted in a 1.0 nm resolution projection map that shows structural elements not revealed with negative staining. Electron microscopic structural data were compared with the crystallographic structure of the 43 kDa N-terminal fragment of the B subunit complexed with a non hydrolysable ATP analogue.
authors.affiliation: Laboratoire de Génétique Moléculaire des Eucaryotes, CNRS, INSERM, Institut de Chimie Biologique, Faculté de Médecine, Strasbourg, France.
authors.forename: H
authors.initials: H
authors.lastname: Celia
authors.forename: L
authors.initials: L
authors.lastname: Hoermann
authors.forename: P
authors.initials: P
authors.lastname: Schultz
authors.forename: L
authors.initials: L
authors.lastname: Lebeau
authors.forename: V
authors.initials: V
authors.lastname: Mallouh
authors.forename: D B
authors.initials: DB
authors.lastname: Wigley
authors.forename: J C
authors.initials: JC
authors.lastname: Wang
authors.forename: C
authors.initials: C
authors.lastname: Mioskowski
authors.forename: P
authors.initials: P
authors.lastname: Oudet
mesh_terms: D001665:Binding Sites; D018360:Crystallography, X-Ray; D027081:DNA Gyrase; D004250:DNA Topoisomerases, Type II / Q000737:chemistry* / Q000648:ultrastructure; D004926:Escherichia coli / Q000737:chemistry / Q000201:enzymology* / Q000648:ultrastructure; D007091:Image Processing, Computer-Assisted; D008854:Microscopy, Electron; D008958:Models, Molecular; D009675:Novobiocin / Q000737:chemistry*; D010743:Phospholipids / Q000737:chemistry*; D014867:Water / Q000737:chemistry
chemical_list: D010743:Phospholipids; D014867:Water; D009675:Novobiocin; D027081:DNA Gyrase; D004250:DNA Topoisomerases, Type II