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The expanding toolkit for structural biology: synchrotrons, X-ray lasers and cryoEM
Structural biology continues to benefit from an expanding toolkit, which is helping to gain unprecedented insight into the assembly and organization of multi-protein machineries, enzyme mechanisms and ligand/inhibitor binding. The combination of results from X-ray free-electron lasers (XFELs), moder...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6400194/ https://www.ncbi.nlm.nih.gov/pubmed/30867914 http://dx.doi.org/10.1107/S2052252519002422 |
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author | Muench, Stephen P. Antonyuk, Svetlana V. Hasnain, S. Samar |
author_facet | Muench, Stephen P. Antonyuk, Svetlana V. Hasnain, S. Samar |
author_sort | Muench, Stephen P. |
collection | PubMed |
description | Structural biology continues to benefit from an expanding toolkit, which is helping to gain unprecedented insight into the assembly and organization of multi-protein machineries, enzyme mechanisms and ligand/inhibitor binding. The combination of results from X-ray free-electron lasers (XFELs), modern synchrotron crystallographic beamlines and cryo-electron microscopy (cryoEM) is proving to be particularly powerful. The highly brilliant undulator beamlines at modern synchrotron facilities have empowered the crystallographic revolution of high-throughput structure determination at high resolution. The brilliance of the X-rays at these crystallographic beamlines has enabled this to be achieved using microcrystals, but at the expense of an increased absorbed X-ray dose and a consequent vulnerability to radiation-induced changes. The advent of serial femtosecond crystallography (SFX) with X-ray free-electron lasers provides a new opportunity in which damage-free structures can be obtained from much smaller crystals (2 µm) and more complex macromolecules, including membrane proteins and multi-protein complexes. For redox enzymes, SFX provides a unique opportunity by providing damage-free structures at both cryogenic and ambient temperatures. The promise of being able to visualize macromolecular structures and complexes at high resolution without the need for crystals using X-rays has remained a dream, but recent technological advancements in cryoEM have made this come true and hardly a month goes by when the structure of a new/novel macromolecular assembly is not revealed. The uniqueness of cryoEM in providing structural information for multi-protein complexes, particularly membrane proteins, has been demonstrated by examples such as respirasomes. The synergistic use of cryoEM and crystallography in lead-compound optimization is highlighted by the example of the visualization of antimalarial compounds in cytochrome bc (1). In this short review, using some recent examples including our own work, we share the excitement of these powerful structural biology methods. |
format | Online Article Text |
id | pubmed-6400194 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-64001942019-03-13 The expanding toolkit for structural biology: synchrotrons, X-ray lasers and cryoEM Muench, Stephen P. Antonyuk, Svetlana V. Hasnain, S. Samar IUCrJ Topical Reviews Structural biology continues to benefit from an expanding toolkit, which is helping to gain unprecedented insight into the assembly and organization of multi-protein machineries, enzyme mechanisms and ligand/inhibitor binding. The combination of results from X-ray free-electron lasers (XFELs), modern synchrotron crystallographic beamlines and cryo-electron microscopy (cryoEM) is proving to be particularly powerful. The highly brilliant undulator beamlines at modern synchrotron facilities have empowered the crystallographic revolution of high-throughput structure determination at high resolution. The brilliance of the X-rays at these crystallographic beamlines has enabled this to be achieved using microcrystals, but at the expense of an increased absorbed X-ray dose and a consequent vulnerability to radiation-induced changes. The advent of serial femtosecond crystallography (SFX) with X-ray free-electron lasers provides a new opportunity in which damage-free structures can be obtained from much smaller crystals (2 µm) and more complex macromolecules, including membrane proteins and multi-protein complexes. For redox enzymes, SFX provides a unique opportunity by providing damage-free structures at both cryogenic and ambient temperatures. The promise of being able to visualize macromolecular structures and complexes at high resolution without the need for crystals using X-rays has remained a dream, but recent technological advancements in cryoEM have made this come true and hardly a month goes by when the structure of a new/novel macromolecular assembly is not revealed. The uniqueness of cryoEM in providing structural information for multi-protein complexes, particularly membrane proteins, has been demonstrated by examples such as respirasomes. The synergistic use of cryoEM and crystallography in lead-compound optimization is highlighted by the example of the visualization of antimalarial compounds in cytochrome bc (1). In this short review, using some recent examples including our own work, we share the excitement of these powerful structural biology methods. International Union of Crystallography 2019-03-01 /pmc/articles/PMC6400194/ /pubmed/30867914 http://dx.doi.org/10.1107/S2052252519002422 Text en © Muench et al. 2019 http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Topical Reviews Muench, Stephen P. Antonyuk, Svetlana V. Hasnain, S. Samar The expanding toolkit for structural biology: synchrotrons, X-ray lasers and cryoEM |
title | The expanding toolkit for structural biology: synchrotrons, X-ray lasers and cryoEM |
title_full | The expanding toolkit for structural biology: synchrotrons, X-ray lasers and cryoEM |
title_fullStr | The expanding toolkit for structural biology: synchrotrons, X-ray lasers and cryoEM |
title_full_unstemmed | The expanding toolkit for structural biology: synchrotrons, X-ray lasers and cryoEM |
title_short | The expanding toolkit for structural biology: synchrotrons, X-ray lasers and cryoEM |
title_sort | expanding toolkit for structural biology: synchrotrons, x-ray lasers and cryoem |
topic | Topical Reviews |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6400194/ https://www.ncbi.nlm.nih.gov/pubmed/30867914 http://dx.doi.org/10.1107/S2052252519002422 |
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