Goniometer-based femtosecond X-ray diffraction of mutant 30S ribosomal subunit crystals

E Han Dao; Raymond G Sierra; Hartawan Laksmono; Henrik T Lemke; Roberto Alonso-Mori; Aaron Coey; Kevin Larsen; Elizabeth L Baxter; Aina E Cohen; S Michael Soltis; Hasan DeMirci

doi:10.1063/1.4919407

Goniometer-based femtosecond X-ray diffraction of mutant 30S ribosomal subunit crystals

Struct Dyn. 2015 Apr 30;2(4):041706. doi: 10.1063/1.4919407. eCollection 2015 Jul.

Authors

Affiliations

¹ Stanford PULSE Institute, SLAC National Accelerator Laboratory , Menlo Park, California 94025, USA.
² Linac Coherent Light Source (LCLS), SLAC National Accelerator Laboratory , Menlo Park, California 94025, USA.
³ Biophysics Program, Stanford University School of Medicine , Stanford, California 94305, USA.
⁴ Stanford Synchrotron Radiation Lightsource (SSRL), SLAC National Accelerator Laboratory , Menlo Park, California 94025, USA.

Abstract

In this work, we collected radiation-damage-free data from a set of cryo-cooled crystals for a novel 30S ribosomal subunit mutant using goniometer-based femtosecond crystallography. Crystal quality assessment for these samples was conducted at the X-ray Pump Probe end-station of the Linac Coherent Light Source (LCLS) using recently introduced goniometer-based instrumentation. These 30S subunit crystals were genetically engineered to omit a 26-residue protein, Thx, which is present in the wild-type Thermus thermophilus 30S ribosomal subunit. We are primarily interested in elucidating the contribution of this ribosomal protein to the overall 30S subunit structure. To assess the viability of this study, femtosecond X-ray diffraction patterns from these crystals were recorded at the LCLS during a protein crystal screening beam time. During our data collection, we successfully observed diffraction from these difficult-to-grow 30S ribosomal subunit crystals. Most of our crystals were found to diffract to low resolution, while one crystal diffracted to 3.2 Å resolution. These data suggest the feasibility of pursuing high-resolution data collection as well as the need to improve sample preparation and handling in order to collect a complete radiation-damage-free data set using an X-ray Free Electron Laser.