title: Serial femtosecond X-ray diffraction of 30S ribosomal subunit microcrystals in liquid suspension at ambient temperature using an X-ray free-electron laser.
abstract: High-resolution ribosome structures determined by X-ray crystallography have provided important insights into the mechanism of translation. Such studies have thus far relied on large ribosome crystals kept at cryogenic temperatures to reduce radiation damage. Here, the application of serial femtosecond X-ray crystallography (SFX) using an X-ray free-electron laser (XFEL) to obtain diffraction data from ribosome microcrystals in liquid suspension at ambient temperature is described. 30S ribosomal subunit microcrystals diffracted to beyond 6 Å resolution, demonstrating the feasibility of using SFX for ribosome structural studies. The ability to collect diffraction data at near-physiological temperatures promises to provide fundamental insights into the structural dynamics of the ribosome and its functional complexes.
authors.affiliation: Molecular Biology, Cell Biology and Biochemistry, Brown University, 185 Meeting Street, Providence, RI 02912, USA. hasan_demirci@brown.edu
authors.forename: Hasan
authors.initials: H
authors.lastname: Demirci
authors.forename: Raymond G
authors.initials: RG
authors.lastname: Sierra
authors.forename: Hartawan
authors.initials: H
authors.lastname: Laksmono
authors.forename: Robert L
authors.initials: RL
authors.lastname: Shoeman
authors.forename: Sabine
authors.initials: S
authors.lastname: Botha
authors.forename: Thomas R M
authors.initials: TR
authors.lastname: Barends
authors.forename: Karol
authors.initials: K
authors.lastname: Nass
authors.forename: Ilme
authors.initials: I
authors.lastname: Schlichting
authors.forename: R Bruce
authors.initials: RB
authors.lastname: Doak
authors.forename: Cornelius
authors.initials: C
authors.lastname: Gati
authors.forename: Garth J
authors.initials: GJ
authors.lastname: Williams
authors.forename: Sébastien
authors.initials: S
authors.lastname: Boutet
authors.forename: Marc
authors.initials: M
authors.lastname: Messerschmidt
authors.forename: Gerwald
authors.initials: G
authors.lastname: Jogl
authors.forename: Albert E
authors.initials: AE
authors.lastname: Dahlberg
authors.forename: Steven T
authors.initials: ST
authors.lastname: Gregory
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Bogan
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D004583:Electrons; D007834:Lasers; D054680:Ribosome Subunits, Small, Bacterial / Q000737:chemistry / Q000648:ultrastructure*; D013696:Temperature; D016963:Thermus thermophilus / Q000737:chemistry*; D014961:X-Ray Diffraction
keywords: 30S ribosomal subunit; X-ray free-electron laser; ribosome; serial femtosecond X-ray crystallography