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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...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Crystallographic Association
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4711619/ https://www.ncbi.nlm.nih.gov/pubmed/26798805 http://dx.doi.org/10.1063/1.4919407 |
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author | Dao, E. Han Sierra, Raymond G. Laksmono, Hartawan Lemke, Henrik T. Alonso-Mori, Roberto Coey, Aaron Larsen, Kevin Baxter, Elizabeth L. Cohen, Aina E. Soltis, S. Michael DeMirci, Hasan |
author_facet | Dao, E. Han Sierra, Raymond G. Laksmono, Hartawan Lemke, Henrik T. Alonso-Mori, Roberto Coey, Aaron Larsen, Kevin Baxter, Elizabeth L. Cohen, Aina E. Soltis, S. Michael DeMirci, Hasan |
author_sort | Dao, E. Han |
collection | PubMed |
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. |
format | Online Article Text |
id | pubmed-4711619 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | American Crystallographic Association |
record_format | MEDLINE/PubMed |
spelling | pubmed-47116192016-01-21 Goniometer-based femtosecond X-ray diffraction of mutant 30S ribosomal subunit crystals Dao, E. Han Sierra, Raymond G. Laksmono, Hartawan Lemke, Henrik T. Alonso-Mori, Roberto Coey, Aaron Larsen, Kevin Baxter, Elizabeth L. Cohen, Aina E. Soltis, S. Michael DeMirci, Hasan Struct Dyn SPECIAL TOPIC: BIOLOGY WITH X-RAY LASERS 2 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. American Crystallographic Association 2015-04-30 /pmc/articles/PMC4711619/ /pubmed/26798805 http://dx.doi.org/10.1063/1.4919407 Text en © 2015 Author(s). 2329-7778/2015/2(4)/041706/10 All article content, except where otherwise noted, is licensed under a Creative Commons Attribution 3.0 Unported License. |
spellingShingle | SPECIAL TOPIC: BIOLOGY WITH X-RAY LASERS 2 Dao, E. Han Sierra, Raymond G. Laksmono, Hartawan Lemke, Henrik T. Alonso-Mori, Roberto Coey, Aaron Larsen, Kevin Baxter, Elizabeth L. Cohen, Aina E. Soltis, S. Michael DeMirci, Hasan Goniometer-based femtosecond X-ray diffraction of mutant 30S ribosomal subunit crystals |
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 |
topic | SPECIAL TOPIC: BIOLOGY WITH X-RAY LASERS 2 |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4711619/ https://www.ncbi.nlm.nih.gov/pubmed/26798805 http://dx.doi.org/10.1063/1.4919407 |
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