5HL4

Acoustic injectors for drop-on-demand serial femtosecond crystallography

Summary for 5HL4

• Entry DOI: 10.2210/pdb5hl4/pdb
• Related: 5F81 5f80
• Descriptor: Ring-hydroxylating dioxygenase, FE2/S2 (INORGANIC) CLUSTER, FE (III) ION, ... (5 entities in total)
• Functional Keywords: ring-hydroxylating dioxygenase, oxidoreductase
• Biological source: Sinorhizobium meliloti 1021 (Ensifer meliloti)
• Total number of polymer chains: 1
• Total formula weight: 46818.24

Authors

Roessler, C.G.,Agarwal, R.,Allaire, M.,Alonso-Mori, R.,Andi, B.,Bachega, J.F.R.,Bommer, M.,Brewster, A.S.,Browne, M.C.,Chatterjee, R.,Cho, E.,Cohen, A.E.,Cowan, M.,Datwani, S.,Davidson, V.L.,Defever, J.,Eaton, B.,Ellson, R.,Feng, Y.,Ghislain, L.P.,Glownia, J.M.,Han, G.,Hattne, J.,Hellmich, J.,Heroux, A.,Ibrahim, M.,Kern, J.,Kuczewski, A.,Lemke, H.T.,Liu, P.,Majlof, L.,McClintock, W.M.,Myers, S.,Nelsen, S.,Olechno, J.,Orville, A.M.,Sauter, N.K.,Soares, A.S.,Soltis, M.S.,Song, H.,Stearns, R.G.,Tran, R.,Tsai, Y.,Uervirojnangkoorn, M.,Wilmot, C.M.,Yachandra, V.,Yano, J.,Yukl, E.T.,Zhu, D.,Zouni, A. (deposition date: 2016-01-14, release date: 2016-01-27, Last modification date: 2024-10-23)

Primary citation

Roessler, C.G.,Agarwal, R.,Allaire, M.,Alonso-Mori, R.,Andi, B.,Bachega, J.F.,Bommer, M.,Brewster, A.S.,Browne, M.C.,Chatterjee, R.,Cho, E.,Cohen, A.E.,Cowan, M.,Datwani, S.,Davidson, V.L.,Defever, J.,Eaton, B.,Ellson, R.,Feng, Y.,Ghislain, L.P.,Glownia, J.M.,Han, G.,Hattne, J.,Hellmich, J.,Heroux, A.,Ibrahim, M.,Kern, J.,Kuczewski, A.,Lemke, H.T.,Liu, P.,Majlof, L.,McClintock, W.M.,Myers, S.,Nelsen, S.,Olechno, J.,Orville, A.M.,Sauter, N.K.,Soares, A.S.,Soltis, S.M.,Song, H.,Stearns, R.G.,Tran, R.,Tsai, Y.,Uervirojnangkoorn, M.,Wilmot, C.M.,Yachandra, V.,Yano, J.,Yukl, E.T.,Zhu, D.,Zouni, A.
Acoustic Injectors for Drop-On-Demand Serial Femtosecond Crystallography.
Structure, 24:631-640, 2016

PubMed Abstract: X-ray free-electron lasers (XFELs) provide very intense X-ray pulses suitable for macromolecular crystallography. Each X-ray pulse typically lasts for tens of femtoseconds and the interval between pulses is many orders of magnitude longer. Here we describe two novel acoustic injection systems that use focused sound waves to eject picoliter to nanoliter crystal-containing droplets out of microplates and into the X-ray pulse from which diffraction data are collected. The on-demand droplet delivery is synchronized to the XFEL pulse scheme, resulting in X-ray pulses intersecting up to 88% of the droplets. We tested several types of samples in a range of crystallization conditions, wherein the overall crystal hit ratio (e.g., fraction of images with observable diffraction patterns) is a function of the microcrystal slurry concentration. We report crystal structures from lysozyme, thermolysin, and stachydrine demethylase (Stc2). Additional samples were screened to demonstrate that these methods can be applied to rare samples.

PubMed: 26996959
DOI: 10.1016/j.str.2016.02.007

Experimental method

X-RAY DIFFRACTION (2.2 Å)