Goniometer-based femtosecond X-ray diffraction of mutant 30S ribosomal subunit crystals
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 L...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
AIP Publishing LLC and ACA
2015-07-01
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| Series: | Structural Dynamics |
| Online Access: | http://dx.doi.org/10.1063/1.4919407 |
| _version_ | 1819232311693017088 |
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| author | 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 |
| author_facet | 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 |
| author_sort | E. Han Dao |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-23T11:58:51Z |
| format | Article |
| id | doaj.art-3523372e8436431785b805c74f048bea |
| institution | Directory Open Access Journal |
| issn | 2329-7778 |
| language | English |
| last_indexed | 2024-12-23T11:58:51Z |
| publishDate | 2015-07-01 |
| publisher | AIP Publishing LLC and ACA |
| record_format | Article |
| series | Structural Dynamics |
| spelling | doaj.art-3523372e8436431785b805c74f048bea2022-12-21T17:48:01ZengAIP Publishing LLC and ACAStructural Dynamics2329-77782015-07-0124041706041706-1010.1063/1.4919407004591SDYGoniometer-based femtosecond X-ray diffraction of mutant 30S ribosomal subunit crystalsE. Han Dao0Raymond G. Sierra1Hartawan Laksmono2Henrik T. Lemke3Roberto Alonso-Mori4Aaron Coey5Kevin Larsen6Elizabeth L. Baxter7Aina E. Cohen8S. Michael Soltis9Hasan DeMirci10 Stanford PULSE Institute, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA Stanford PULSE Institute, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA Stanford PULSE Institute, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA Linac Coherent Light Source (LCLS), SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA Linac Coherent Light Source (LCLS), SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA Biophysics Program, Stanford University School of Medicine, Stanford, California 94305, USA Biophysics Program, Stanford University School of Medicine, Stanford, California 94305, USA Stanford Synchrotron Radiation Lightsource (SSRL), SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA Stanford Synchrotron Radiation Lightsource (SSRL), SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA Stanford Synchrotron Radiation Lightsource (SSRL), SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA Stanford PULSE Institute, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USAIn 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.http://dx.doi.org/10.1063/1.4919407 |
| spellingShingle | 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 Goniometer-based femtosecond X-ray diffraction of mutant 30S ribosomal subunit crystals Structural Dynamics |
| title | Goniometer-based femtosecond X-ray diffraction of mutant 30S ribosomal subunit crystals |
| title_full | Goniometer-based femtosecond X-ray diffraction of mutant 30S ribosomal subunit crystals |
| title_fullStr | Goniometer-based femtosecond X-ray diffraction of mutant 30S ribosomal subunit crystals |
| title_full_unstemmed | Goniometer-based femtosecond X-ray diffraction of mutant 30S ribosomal subunit crystals |
| title_short | Goniometer-based femtosecond X-ray diffraction of mutant 30S ribosomal subunit crystals |
| title_sort | goniometer based femtosecond x ray diffraction of mutant 30s ribosomal subunit crystals |
| url | http://dx.doi.org/10.1063/1.4919407 |
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