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Segmented flow generator for serial crystallography at the European X-ray free electron laser
Serial femtosecond crystallography (SFX) with X-ray free electron lasers (XFELs) allows structure determination of membrane proteins and time-resolved crystallography. Common liquid sample delivery continuously jets the protein crystal suspension into the path of the XFEL, wasting a vast amount of s...
| Autores principales: | , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , |
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| Formato: | Online Artículo Texto |
| Lenguaje: | English |
| Publicado: |
Nature Publishing Group UK
2020
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| Materias: | |
| Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7481229/ https://www.ncbi.nlm.nih.gov/pubmed/32908128 http://dx.doi.org/10.1038/s41467-020-18156-7 |
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| author | Echelmeier, Austin Cruz Villarreal, Jorvani Messerschmidt, Marc Kim, Daihyun Coe, Jesse D. Thifault, Darren Botha, Sabine Egatz-Gomez, Ana Gandhi, Sahir Brehm, Gerrit Conrad, Chelsie E. Hansen, Debra T. Madsen, Caleb Bajt, Saša Meza-Aguilar, J. Domingo Oberthür, Dominik Wiedorn, Max O. Fleckenstein, Holger Mendez, Derek Knoška, Juraj Martin-Garcia, Jose M. Hu, Hao Lisova, Stella Allahgholi, Aschkan Gevorkov, Yaroslav Ayyer, Kartik Aplin, Steve Ginn, Helen Mary Graafsma, Heinz Morgan, Andrew J. Greiffenberg, Dominic Klujev, Alexander Laurus, Torsten Poehlsen, Jennifer Trunk, Ulrich Mezza, Davide Schmidt, Bernd Kuhn, Manuela Fromme, Raimund Sztuk-Dambietz, Jolanta Raab, Natascha Hauf, Steffen Silenzi, Alessandro Michelat, Thomas Xu, Chen Danilevski, Cyril Parenti, Andrea Mekinda, Leonce Weinhausen, Britta Mills, Grant Vagovic, Patrik Kim, Yoonhee Kirkwood, Henry Bean, Richard Bielecki, Johan Stern, Stephan Giewekemeyer, Klaus Round, Adam R. Schulz, Joachim Dörner, Katerina Grant, Thomas D. Mariani, Valerio Barty, Anton Mancuso, Adrian P. Weierstall, Uwe Spence, John C. H. Chapman, Henry N. Zatsepin, Nadia Fromme, Petra Kirian, Richard A. Ros, Alexandra |
| author_facet | Echelmeier, Austin Cruz Villarreal, Jorvani Messerschmidt, Marc Kim, Daihyun Coe, Jesse D. Thifault, Darren Botha, Sabine Egatz-Gomez, Ana Gandhi, Sahir Brehm, Gerrit Conrad, Chelsie E. Hansen, Debra T. Madsen, Caleb Bajt, Saša Meza-Aguilar, J. Domingo Oberthür, Dominik Wiedorn, Max O. Fleckenstein, Holger Mendez, Derek Knoška, Juraj Martin-Garcia, Jose M. Hu, Hao Lisova, Stella Allahgholi, Aschkan Gevorkov, Yaroslav Ayyer, Kartik Aplin, Steve Ginn, Helen Mary Graafsma, Heinz Morgan, Andrew J. Greiffenberg, Dominic Klujev, Alexander Laurus, Torsten Poehlsen, Jennifer Trunk, Ulrich Mezza, Davide Schmidt, Bernd Kuhn, Manuela Fromme, Raimund Sztuk-Dambietz, Jolanta Raab, Natascha Hauf, Steffen Silenzi, Alessandro Michelat, Thomas Xu, Chen Danilevski, Cyril Parenti, Andrea Mekinda, Leonce Weinhausen, Britta Mills, Grant Vagovic, Patrik Kim, Yoonhee Kirkwood, Henry Bean, Richard Bielecki, Johan Stern, Stephan Giewekemeyer, Klaus Round, Adam R. Schulz, Joachim Dörner, Katerina Grant, Thomas D. Mariani, Valerio Barty, Anton Mancuso, Adrian P. Weierstall, Uwe Spence, John C. H. Chapman, Henry N. Zatsepin, Nadia Fromme, Petra Kirian, Richard A. Ros, Alexandra |
| author_sort | Echelmeier, Austin |
| collection | PubMed |
| description | Serial femtosecond crystallography (SFX) with X-ray free electron lasers (XFELs) allows structure determination of membrane proteins and time-resolved crystallography. Common liquid sample delivery continuously jets the protein crystal suspension into the path of the XFEL, wasting a vast amount of sample due to the pulsed nature of all current XFEL sources. The European XFEL (EuXFEL) delivers femtosecond (fs) X-ray pulses in trains spaced 100 ms apart whereas pulses within trains are currently separated by 889 ns. Therefore, continuous sample delivery via fast jets wastes >99% of sample. Here, we introduce a microfluidic device delivering crystal laden droplets segmented with an immiscible oil reducing sample waste and demonstrate droplet injection at the EuXFEL compatible with high pressure liquid delivery of an SFX experiment. While achieving ~60% reduction in sample waste, we determine the structure of the enzyme 3-deoxy-D-manno-octulosonate-8-phosphate synthase from microcrystals delivered in droplets revealing distinct structural features not previously reported. |
| format | Online Article Text |
| id | pubmed-7481229 |
| institution | National Center for Biotechnology Information |
| language | English |
| publishDate | 2020 |
| publisher | Nature Publishing Group UK |
| record_format | MEDLINE/PubMed |
| spelling | pubmed-74812292020-09-21 Segmented flow generator for serial crystallography at the European X-ray free electron laser Echelmeier, Austin Cruz Villarreal, Jorvani Messerschmidt, Marc Kim, Daihyun Coe, Jesse D. Thifault, Darren Botha, Sabine Egatz-Gomez, Ana Gandhi, Sahir Brehm, Gerrit Conrad, Chelsie E. Hansen, Debra T. Madsen, Caleb Bajt, Saša Meza-Aguilar, J. Domingo Oberthür, Dominik Wiedorn, Max O. Fleckenstein, Holger Mendez, Derek Knoška, Juraj Martin-Garcia, Jose M. Hu, Hao Lisova, Stella Allahgholi, Aschkan Gevorkov, Yaroslav Ayyer, Kartik Aplin, Steve Ginn, Helen Mary Graafsma, Heinz Morgan, Andrew J. Greiffenberg, Dominic Klujev, Alexander Laurus, Torsten Poehlsen, Jennifer Trunk, Ulrich Mezza, Davide Schmidt, Bernd Kuhn, Manuela Fromme, Raimund Sztuk-Dambietz, Jolanta Raab, Natascha Hauf, Steffen Silenzi, Alessandro Michelat, Thomas Xu, Chen Danilevski, Cyril Parenti, Andrea Mekinda, Leonce Weinhausen, Britta Mills, Grant Vagovic, Patrik Kim, Yoonhee Kirkwood, Henry Bean, Richard Bielecki, Johan Stern, Stephan Giewekemeyer, Klaus Round, Adam R. Schulz, Joachim Dörner, Katerina Grant, Thomas D. Mariani, Valerio Barty, Anton Mancuso, Adrian P. Weierstall, Uwe Spence, John C. H. Chapman, Henry N. Zatsepin, Nadia Fromme, Petra Kirian, Richard A. Ros, Alexandra Nat Commun Article Serial femtosecond crystallography (SFX) with X-ray free electron lasers (XFELs) allows structure determination of membrane proteins and time-resolved crystallography. Common liquid sample delivery continuously jets the protein crystal suspension into the path of the XFEL, wasting a vast amount of sample due to the pulsed nature of all current XFEL sources. The European XFEL (EuXFEL) delivers femtosecond (fs) X-ray pulses in trains spaced 100 ms apart whereas pulses within trains are currently separated by 889 ns. Therefore, continuous sample delivery via fast jets wastes >99% of sample. Here, we introduce a microfluidic device delivering crystal laden droplets segmented with an immiscible oil reducing sample waste and demonstrate droplet injection at the EuXFEL compatible with high pressure liquid delivery of an SFX experiment. While achieving ~60% reduction in sample waste, we determine the structure of the enzyme 3-deoxy-D-manno-octulosonate-8-phosphate synthase from microcrystals delivered in droplets revealing distinct structural features not previously reported. Nature Publishing Group UK 2020-09-09 /pmc/articles/PMC7481229/ /pubmed/32908128 http://dx.doi.org/10.1038/s41467-020-18156-7 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
| spellingShingle | Article Echelmeier, Austin Cruz Villarreal, Jorvani Messerschmidt, Marc Kim, Daihyun Coe, Jesse D. Thifault, Darren Botha, Sabine Egatz-Gomez, Ana Gandhi, Sahir Brehm, Gerrit Conrad, Chelsie E. Hansen, Debra T. Madsen, Caleb Bajt, Saša Meza-Aguilar, J. Domingo Oberthür, Dominik Wiedorn, Max O. Fleckenstein, Holger Mendez, Derek Knoška, Juraj Martin-Garcia, Jose M. Hu, Hao Lisova, Stella Allahgholi, Aschkan Gevorkov, Yaroslav Ayyer, Kartik Aplin, Steve Ginn, Helen Mary Graafsma, Heinz Morgan, Andrew J. Greiffenberg, Dominic Klujev, Alexander Laurus, Torsten Poehlsen, Jennifer Trunk, Ulrich Mezza, Davide Schmidt, Bernd Kuhn, Manuela Fromme, Raimund Sztuk-Dambietz, Jolanta Raab, Natascha Hauf, Steffen Silenzi, Alessandro Michelat, Thomas Xu, Chen Danilevski, Cyril Parenti, Andrea Mekinda, Leonce Weinhausen, Britta Mills, Grant Vagovic, Patrik Kim, Yoonhee Kirkwood, Henry Bean, Richard Bielecki, Johan Stern, Stephan Giewekemeyer, Klaus Round, Adam R. Schulz, Joachim Dörner, Katerina Grant, Thomas D. Mariani, Valerio Barty, Anton Mancuso, Adrian P. Weierstall, Uwe Spence, John C. H. Chapman, Henry N. Zatsepin, Nadia Fromme, Petra Kirian, Richard A. Ros, Alexandra Segmented flow generator for serial crystallography at the European X-ray free electron laser |
| title | Segmented flow generator for serial crystallography at the European X-ray free electron laser |
| title_full | Segmented flow generator for serial crystallography at the European X-ray free electron laser |
| title_fullStr | Segmented flow generator for serial crystallography at the European X-ray free electron laser |
| title_full_unstemmed | Segmented flow generator for serial crystallography at the European X-ray free electron laser |
| title_short | Segmented flow generator for serial crystallography at the European X-ray free electron laser |
| title_sort | segmented flow generator for serial crystallography at the european x-ray free electron laser |
| topic | Article |
| url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7481229/ https://www.ncbi.nlm.nih.gov/pubmed/32908128 http://dx.doi.org/10.1038/s41467-020-18156-7 |
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