Segmented flow generator for serial crystallography at the European X-ray free electron laser
-
Published:2020-09-09
Issue:1
Volume:11
Page:
-
ISSN:2041-1723
-
Container-title:Nature Communications
-
language:en
-
Short-container-title:Nat Commun
Author:
Echelmeier AustinORCID, Cruz Villarreal JorvaniORCID, Messerschmidt Marc, Kim Daihyun, Coe Jesse D.ORCID, Thifault DarrenORCID, Botha Sabine, Egatz-Gomez AnaORCID, Gandhi Sahir, Brehm Gerrit, Conrad Chelsie E., Hansen Debra T.ORCID, Madsen Caleb, Bajt SašaORCID, Meza-Aguilar J. Domingo, Oberthür DominikORCID, Wiedorn Max O., Fleckenstein Holger, Mendez Derek, Knoška JurajORCID, Martin-Garcia Jose M.ORCID, Hu Hao, Lisova StellaORCID, Allahgholi Aschkan, Gevorkov YaroslavORCID, Ayyer KartikORCID, Aplin Steve, Ginn Helen MaryORCID, Graafsma Heinz, Morgan Andrew J., Greiffenberg Dominic, Klujev AlexanderORCID, Laurus Torsten, Poehlsen JenniferORCID, Trunk UlrichORCID, Mezza DavideORCID, Schmidt BerndORCID, Kuhn Manuela, Fromme RaimundORCID, Sztuk-Dambietz Jolanta, Raab Natascha, Hauf Steffen, Silenzi Alessandro, Michelat ThomasORCID, Xu Chen, Danilevski Cyril, Parenti Andrea, Mekinda Leonce, Weinhausen Britta, Mills Grant, Vagovic Patrik, Kim Yoonhee, Kirkwood HenryORCID, Bean Richard, Bielecki Johan, Stern Stephan, Giewekemeyer Klaus, Round Adam R., Schulz JoachimORCID, Dörner Katerina, Grant Thomas D.ORCID, Mariani Valerio, Barty Anton, Mancuso Adrian P., Weierstall Uwe, Spence John C. H., Chapman Henry N.ORCID, Zatsepin NadiaORCID, Fromme Petra, Kirian Richard A., Ros AlexandraORCID
Abstract
AbstractSerial 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.
Funder
National Science Foundation U.S. Department of Health & Human Services | National Institutes of Health Deutsche Forschungsgemeinschaft
Publisher
Springer Science and Business Media LLC
Subject
General Physics and Astronomy,General Biochemistry, Genetics and Molecular Biology,General Chemistry
Reference84 articles.
1. Bowman, S. E., Bridwell-Rabb, J. & Drennan, C. L. Metalloprotein crystallography: more than a structure. Acc. Chem. Res. 49, 695–702 (2016). 2. Chapman, H. N. et al. Femtosecond X-ray protein nanocrystallography. Nature 470, 73–77 (2011). 3. Andersson, R. et al. Serial femtosecond crystallography structure of cytochrome c oxidase at room temperature. Sci. Rep. 7, 4518 (2017). 4. Pande, K. et al. Femtosecond structural dynamics drives the trans/cis isomerization in photoactive yellow protein. Science 352, 725–729 (2016). 5. Coquelle, N. et al. Chromophore twisting in the excited state of a photoswitchable fluorescent protein captured by time-resolved serial femtosecond crystallography. Nat. Chem. 10, 31–37 (2018).
Cited by
32 articles.
订阅此论文施引文献
订阅此论文施引文献，注册后可以免费订阅5篇论文的施引文献，订阅后可以查看论文全部施引文献
|
|