Allen Orville


Allen is the Principal Scientist of the UK XFEL Hub. Dr. Orville received his BS and Ph.D. degrees from the University of Minnesota (Minneapolis/St. Paul), did his post-doctoral training with the Howard Hughes Medical Institute at the University of Oregon (Eugene, OR), was an Assistant Professor at the Georgia Institute of Technology (Atlanta, GA), and was awarded tenure as a Biophysicist in the Biology Department and the Photon Sciences Division at Brookhaven National Laboratory (Upton, NY). In October 2015 he joined Diamond Light Source as group leader of the new XFEL Hub.

Email: [email protected]
Tel: +44 (0) 1235 567505

Macromolecular crystallography and time-resolved structural biology

Microspectroscopy correlated with x-ray crystallography

Serial crystallography at synchrotrons and XFELs

Current Research Interests

Despite their immense value, the atomic coordinates derived from ground-state x-ray crystal structures reveal only a portion of the functional story for many important and complex systems in Biology. Consequently, a frontier challenge for many structural biologists is to determine time-resolved crystal structures directly from systems engaged in catalysis. In structural biology this leads to a desire to collect structural, spectroscopic, and if possible, kinetic data too, from the same crystalline samples. To these ends, single-crystal electronic absorption and Raman spectroscopies can be correlated with x-ray crystal structures at several synchrotron facilities, including Diamond Light Source and the Central Laser Facility. However, a set of general tools to fully correlate enzyme kinetics with time-resolved crystallography are not yet readily available. To address this grand challenge, we are working to extended enzyme kinetics strategies to crystalline samples, and to couple these methods to room-temperature, time-resolved, serial crystallographic methods with correlations to single-crystal spectroscopy and other structural methods. Achieving our goals will be transformative and impact nearly all aspects of structural biology.

Serial crystallography is a rapidly developing field with the potential to produce time-resolved, atomic resolution movies of macromolecules engaged in catalysis. It uses a large number of several µm3 or smaller crystals, from each of which only a small fraction of diffraction data is collected, which are then merged together to form the whole dataset. Our guiding hypothesis is that most enzyme microcrystals (on the order 2 μm3 and smaller) will equilibrate with substrate(s) many times faster than their overall reaction cycle. If this technique is applied to µm3-sized crystals at synchrotrons, then time domains in the µs and longer can be probed. X-ray free electron lasers (XFEL) sources enable us to collect diffraction and spectroscopic data from the same sample on the fs and longer time-scale and with smaller crystals.

We are developing tools that will exploit a variety of triggering strategies for use at i) the LCLS at SLAC National Accelerator Laboratory in the USA, ii) SACLA in Japan, iii) on the Harwell campus at Diamond Light Source, the Central Laser Facility, and for cryo-EM at the electron Bio-Imaging Centre, as well as iv) at the European XFEL in Hamburg, Germany. For example, our novel LCLS acoustic injection systems use focused sound waves to eject on demand picoliter to nanoliter crystal-containing droplets directly into the LCLS x-ray pulse. Our acoustic conveyor belt moves droplets through a series of visible-light laser beams to support time-resolved, pump-probe studies. At the LCLS we are studying several light-triggered reactions, including phytochromes (photon receptors that impact optogentics) and photosystem II (spectroscopy and structure of the water oxidizing complex). Rapid mixing jets and microfluidics systems will support enzyme turnover in time-resolved serial crystallography, which will focus initially on metalloenzyme catalysis.


Bradly Davy, Danny Axford, John H. Beale, Agata Butryn, Peter Docker, Ali Ebrahim, Gabriel Leen, Allen M. Orville, Robin L. Owen and Pierre Aller, “Reducing sample consumption for serial crystallography using acoustic drop ejection” Journal of Synchrotron Radiation (2019) 26 (in press),
Jan Kern, Ruchira Chatterjee, Iris D. Young, Franklin D. Fuller, Louise Lassalle, Mohamed Ibrahim, Sheraz Gul, Thomas Fransson, Aaron S. Brewster, Roberto Alonso-Mori4, Rana Hussein, Miao Zhang, Lacey Douthit, Casper de Lichtenberg, Mun Hon Cheah, Dmitry Shevela, Julia Wersig, Ina Seufert, Dimosthenis Sokaras, Ernest Pastor, Clemens Weninger, Thomas Kroll, Raymond G. Sierra, Pierre Aller, Agata Butryn, Allen M. Orville, Mengning Liang, Alexander Batyuk, Jason E. Koglin, Sergio Carbajo, Sébastien Boutet, Nigel W. Moriarty, James M. Holton, Holger Dobbek, Paul D. Adams, Uwe Bergmann, Nicholas K. Sauter, Athina Zouni*, Johannes Messinger*, Junko Yano*, Vittal K. Yachandra*, “Structures of the intermediates of Kok’s photosynthetic water oxidation clock” Nature(2018) 563: 421-425; PMCID: PMC6485242; DOI: 10.1038/s41586-018-0681-2
Max O. Wiedorn, Dominik Oberthür, Richard Bean, Robin Schubert, Nadine Werner, Brian Abbey, Martin Aepfelbacher, Luigi Adriano, Aschkan Allahgholi, Nasser Al-Qudami, Jakob Andreasson, Steve Aplin, Salah Awel, Kartik Ayyer, Saša Bajt, Imrich Barák, Sadia Bari, Johan Bielecki, Sabine Botha, Djelloul Boukhelef, Wolfgang Brehm, Sandor Brockhauser, Igor Cheviakov, Matthew A. Coleman, Francisco Cruz-Mazo, Cyril Danilevski, Connie Darmanin, R. Bruce Doak, Martin Domaracky, Katerina Dörner, Yang Du, Hans Fangohr, Holger Fleckenstein, Matthias Frank, Petra Fromme, Alfonso M. Gañán-Calvo, Yaroslav Gevorkov, Klaus Giewekemeyer, Helen Mary Ginn, Heinz Graafsma, Rita Graceffa, Dominic Greiffenberg, Lars Gumprecht, Peter Göttlicher, Janos Hajdu, Steffen Hauf, Michael Heymann, Susannah Holmes, Daniel Horke, Mark S. Hunter, Siegfried Imlau, Alexander Kaukher, Yoonhee Kim, Alexander Klyuev, Juraj Knoška, Bostjan Kobe, Manuela Kuhn, Christopher Kupitz, Jochen Kupper, Janine Mia Lahey-Rudolph, Torsten Laurus, Karoline Le Cong, Romain Letrun, P. Lourdu Xavier, Luis Maia, Filipe R.N.C. Maia, Valerio Mariani, Marc Messerschmidt, Markus Metz, Davide Mezza, Thomas Michelat, Grant Mills, Diana C.F. Monteiro, Andrew Morgan, Kerstin Muhlig, Anna Munke, Astrid Münnich, Julia Nette, Keith A. Nugent, Theresa Nuguid, Allen M. Orville, Suraj Pandey, Gisel Pena, Pablo Villanueva-Perez, Jennifer Poehlsen, Gianpietro Previtali, Lars Redecke, Winnie Maria Riekehr, Holger Rohde, Adam Round, Tatiana Safenreiter, Iosifina Sarrou, Tokushi Sato, Marius Schmidt, Bernd Schmitt, Robert Schönherr, Joachim Schulz, Jonas A. Sellberg, M. Marvin Seibert, Carolin Seuring, Megan L. Shelby, Robert L. Shoeman, Marcin Sikorski, Alessandro Silenzi, Claudiu A. Stan, Xintian Shi, Stephan Stern, Jola Sztuk-Dambietz, Janusz Szuba, Aleksandra Tolstikova, Martin Trebbin, Ulrich Trunk, Patrick Vagovic, Thomas Ve, Britta Weinhausen, Thomas A. White, Krzysztof Wrona, Chen Xu, Oleksandr Yefanov, Nadia Zatsepin, Jiaguo Zhang, Markus Perbandt, Adrian P. Mancuso, Christian Betzel, Henry Chapman*, & Anton Barty*, “Megahertz serial crystallography” Nature Communications (2018) 9, 4025  doi: 10.1038/s41467-018-06156-7; PMCID: PMC6168542
Orville, A. M. “Entering an era of dynamic structural biology”, BMC Biol (2018) 16, 55 doi: 10.1186/s12915-018-0533-4; PMCID: PMC5977480; DOI: 10.1186/s12915-018-0533-4
Christian G. Burton, Danny Axford, Andrew M. J. Edwards, Richard Gildea, Rob H. Morris, Michael I. Newton,Allen M. Orville, Mark Prince, Paul D. Topham, and Peter T. Docker “An Acoustic On-Chip Goniometer for Room Temperature Macromolecular Crystallography” Lab on a Chip (2017) 17, 4225-4230; doi: 10.1039/c7lc00812k; PMID: 29124258
Jonathan Mark Grimes, David Hall, Alun Ashton, Gwyndaf Evans, Robin Owen, Armin Wagner, Katherine McAuley, Frank von Delft, Allen M. Orville, Thomas Sorensen, Martin Walsh, Helen Mary Ginn, and David Ian Stuart, “Where is crystallography going?” Acta Crystallographica Section D: Structural Biology (2018) 74: 152-166; PMCID: PMC5947779
Allen M. Orville, “The XFEL Hub at Diamond: A Bridge between Structural Biologists and Facilities” (2017)Synchrotron Radiation News, Vol. 30, 37 – 39;
Peter Docker, Robert Morris, Michael Newton, Elizabeth Dye, James Kay, John Beale, Danny Axford, Allen Orville, Dave Stuart and Gabriel Leen, “Pico-litre Sample Introduction and Acoustic Levitation Systems for Time Resolved Protein Crystallography Experiments at XFELs” (2017) Sensors & Transducers, Vol. 214, pp. 39 – 45.
Franklin D. Fuller, Sheraz Gul, Ruchira Chatterjee, Ernest S. Burgie, Iris D. Young, Hugo Lebrette, Vivek Srinivas, Aaron S. Brewster, Tara Michels-Clark, Jonathan A. Clinger, Babak Andi, Mohamed Ibrahim, Ernest Pastor, Casper de Lichtenberg, Rana Hussein, Christopher J. Pollock, Miao Zhang, Claudiu A. Stan, Thomas Kroll, Thomas Fransson, Clemens Weninger, Markus Kubin, Pierre Aller, Louise Lassalle, Philipp Bräuer, Muhamed Amin, Sergey Koroidov, Christian G. Roessler, Marc Allaire, Raymond G. Sierra, Peter T. Docker, James M. Glownia, Silke Nelson, Jason Koglin, Diling Zhu, Matthieu Chollet, Sanghoon Song, Henrik Lemke, Mengning Liang, Dimosthenis Sokaras, Roberto Alonso-Mori, Athina Zouni, Johannes Messinger, Uwe Bergmann, Amie K. Boal, J. Martin Bollinger, Jr., Carsten Krebs, Martin Högbom, George N. Phillips, Jr., Richard D. Vierstra, Nicholas K. Sauter, AllenM. Orville, Jan Kern, Vittal K. Yachandra, Junko Yano, “Drop-on-Demand Sample Delivery for Studying Biocatalysts in Action at XFELs” (2017) Nature Methods 14, 443–449; doi:10.1038/nmeth.4195; PMCID: PMC5376230
Allen M. Orville “Acoustic Methods for On-Demand Sample Injection into XFEL Beams” in X-Ray Free Electron Lasers: Applications in Materials, Chemistry and Biology edited by Uwe Bergmann, Junko Yano and Vittal Yachandra, published by the Royal Society of Chemistry, Cambridge UK (2017) pp 348 – 363 
Iris D. Young, Mohamed Ibrahim, Ruchira Chatterjee, Sheraz Gul, Franklin D. Fuller, Sergey Koroidov, Aaron S. Brewster, Rosalie Tran, Roberto Alonso-Mori, Thomas Kroll, Tara Michels-Clark, Hartawan Laksmono, Raymond G. Sierra, Claudiu A. Stan, Rana Hussein, Miao Zhang, Lacey Douthit, Markus Kubin, Casper de Lichtenberg, Long Vo Pham, Håkan Nilsson, Mun Hon Cheah, Dmitriy Shevela, Claudio Saracini, Mackenzie A. Bean, Ina Seuffert, Dimosthenis Sokaras, Tsu-Chien Weng, Ernest Pastor, Clemens Weninger, Thomas Fransson, Louise Lassalle, Philipp Bräuer, Pierre Aller, Peter T. Docker, Babak Andi, Allen M. Orville, James M. Glownia, Silke Nelson, Marcin Sikorski, Diling Zhu, Mark S. Hunter, Thomas J. Lane, Andy Aquila, Jason E. Koglin, Joseph Robinson, Mengning Liang, Sébastien Boutet, Artem Y. Lyubimov, Monarin Uervirojnangkoorn, Nigel W. Moriarty, Dorothee Liebschner, Pavel V. Afonine, David G. Waterman, Gwyndaf Evans, Philippe Wernet, Holger Dobbek, William I. Weis, Axel T. Brunger, Petrus H. Zwart, Paul D. Adams, Athina Zouni, Johannes Messinger, Uwe Bergmann, Nicholas K. Sauter, Jan Kern, Vittal K. Yachandra & Junko Yano, “Structure of photosystem II and substrate binding at room temperature” (2016) Nature 540, 453–457; doi:10.1038/nature20161; PMCID: PMC5201176
Christian G. Roessler, Rakhi Agarwal, Marc Allaire*, Roberto Alonso-Mori, Babak Andi, José F. R. Bachega, Martin Bommer, Aaron S. Brewster, Michael C. Browne, Ruchira Chatterjee, Eunsun Cho, Aina E. Cohen, Matthew Cowan, Sammy Datwani, Victor L. Davidson, Jim Defever, Brent Eaton, Richard Ellson, Yiping Feng, Lucien P. Ghislain, James M. Glownia, Guangye Han, Johan Hattne, Julia Hellmich, Annie Héroux, Mohamed Ibrahim, Jan Kern, Anthony Kuczewski, Henrik T. Lemke, Pinghua Liu, Lars Majlof, William M. McClintock, Stuart Myers, Silke Nelsen, Joe Olechno, Allen M. Orville*, Nicholas K. Sauter, Alexei S. Soares*, S. Michael Soltis, Heng Song,  Richard G. Stearns, Rosalie Tran, Yingssu Tsai, Monarin Uervirojnangkoorn, Carrie M. Wilmot, Vittal Yachandra, Junko Yano, Erik T. Yukl, Diling Zhu, Athina Zouni; “Acoustic injectors for drop-on-demand serial femtosecond crystallography” (2016) Structure 24, 631-640; PMID: 26996959; PMCID: PMC4920001

Feifei Li, E. Sethe Burgie, Tao Yu, Annie Héroux, George C. Schatz, Richard D. Vierstra, Allen M. Orville, "X‑ray Radiation Induces Deprotonation of the Bilin Chromophore in Crystalline D. radiodurans Phytochrome", J. Am. Chem. Soc. (2015) 137, 2792-2795; DOI: 10.1021/ja510923m; PMID: 25650486

Lu Huo, Ian Davis, Fange Liu, Babak Andi, Shingo Esaki, Hiroaki Iwaki, Yoshie Hasegawa, Allen M. Orville Nature Communications (2015) Jan 7; 6:5935; DOI: 10.1038/ncomms6935; PMID: 25565451

Héroux, A., Allaire, M., Buono, R., Cowan, M. L., Dvorak, J. Flaks, L. LaMarra, S., Myers, S. F., Orville, A. M., Robinson, H. H., Roessler, C. G., Schneider, D. K., Shea-McCarthy, G., Skinner, J. M., Skinner, M., Soares, A. S., Sweet, R. M. and Berman, L. E. "Macromolecular Crystallography beamline X25 at the NSLS" J. Synchrotron Rad. (2014). 21, 627-632; PMID: 24763654

Roessler, C. G., Kuczewski, A., Orville, A. M., Allaire, M., Soares, A. S., Héroux, A., "Acoustic Methods for High-Throughput Protein Crystal Mounting at Next-Generation Macromolecular Crystallographic Beamlines" J. Synchrotron Radiation 20, 805 - 808 (2013) PMID: 23955046; please see the associated video at:

Daughtry, K.D., Xiao, Y., Stoner-Ma, D., Cho, E., Orville, A.M.*, Liu, P.*, and Allen, K.N.*, "Quaternary Ammonium Oxidative Demethylation: X-ray Crystallographic, Resonance Raman and UV-visible Spectroscopic Analysis of a Rieske-type Demethylase," J. Am. Chem. Soc., 134 (5), 2823 – 2834 (2012) PMID: 22224443

Orville, A.M."Single-Crystal Spectroscopy Correlated with X-Ray Crystallography Provides Complementary Perspectives on Macromolecular Function" Chapter 9 in Methods in Protein Biochemistry, edited by Harald Tschesche, ISBN 978-3-11-218934-4, DE GRUYTER, Berlin, Germany, pages 143-164 (2012)

Soares, A.S., Engel, M.A., Stearns, R., Datwani, S., Olechno, J., Ellson, R., Skinner, J.M., Allaire, M., and Orville, A.M. "Acoustically Mounted Microcrystals Yield High Resolution X-ray Structures" a Rapid Report in Biochemistry 50, 4399-4401 (2011); PMID: 21542590

Orville, A.M., Buono, R., Cowan, M., Héroux, A., Schneider, D.K., Shea-McCarthy, G., Skinner, J.M., Skinner, M.J., Stoner-Ma, D., Sweet, R.M. "Correlated Single-Crystal Electron Absorption Spectroscopy and X-ray Crystallography at NSLS Beamline X26-C" J. Synchrotron Radiation 18, 358-366 (2011); PMID: 21525643

Stoner-Ma, D., Skinner, J.M., Héroux, A., Schneider, D.K., Sweet, R.M. Orville, A.M. "Single-Crystal Raman Spectroscopy and X-ray Crystallography at Beamline X26-C of the NSLS" J. Synchrotron Rad. 18, 37-40 (2011); PMID: 21169688

Finnegan, S., Yuan, H., Wang, Y-F., Orville, A.M., Weber, I.T., Gadda, G. "Structural and Kinetic Studies on the Ser101Ala Variant of Choline Oxidase" Archives of Biochemistry and Biophysics 501, 207-213 (2010); PMID: 20561507

Yi, J., Orville, A.M., Skinner, J.M., Skinner, M.J., Richter-Addo, G.B. "Synchrotron X-ray-Induced Photoreduction of Ferric Myoglobin Nitrite Crystals Gives the Ferrous Derivative with Retention of the O-bonded Nitrite Ligand" Rapid Report in Biochemistry, 49, 5969-5971 (2010). PMID: 20568729

Major, D.T., Héroux, A., Orville, A.M., Valley, M.P., Fitzpatrick P.F., Gao, J. "Differential quantum tunneling contributions in nitroalkane oxidase catalyzed and the uncatalyzed proton transfer reaction" Proc. Nat. Acad. Sci. USA, 106, 20734-20739 (2009); PMID: 19926855

Héroux, A., Bozinovski, D.M., Valley, M.P., Fitzpatrick, P.F. and Orville, A.M. "Crystal Structures of Intermediates in the Nitroalkane Oxidase Reaction", Biochemistry 48, 3407-3416 (2009); PMID: 19265437

Orville, A.M., Lountos, G.T., Finne­gan, S., Gadda, G., Prabhakar R. "Crystallographic, Spectroscopic, and Computational Analysis of a Flavin-C4a-Oxygen Adduct in Choline Oxidase" Biochemistry 48, 720-728 (2009); PMID: 19133805

Quaye, O., Lountos, G.T., Fan, F., Orville, A.M., Gadda, G. "Role of Glu312 in Binding and Positioning of the Substrate for the Hydride Transfer Reaction in Choline Oxidase" Biochemistry 47, 243-256 (2008); PMID: 18072756

Fitzpatrick, P.F., Bozinovski, D.M., Héroux, A., Shaw, P.G., Valley, M.P., Orville, A.M., "Mechanistic and Structural Analyses of the Roles of Arg409 and Asp402 in the Reaction of the Flavoprotein Nitroalkane Oxidase", Biochemistry 46, 13800-13808 (2007)

Pau, M.Y.M., Davis, M.I., Orville, A.M., Lipscomb, J.D. and Solomon, E.I.; "Spectroscopic and Electronic Structure Study of the Enzyme-Substrate Complex of Intradiol Dioxygenases: Substrate Activation by a High-Spin Ferric Non-heme Iron Site" J. Am. Chem. Soc. 129, 1944-1958 (2007)

Lountos, G.T., Jiang, R., Wellborn, W.B., Thaler, T.L., Bommarius, A.S. and Orville, A.M., "The Crystal Structure of NAD(P)H Oxidase from Lactobacillus sanfranciscensis: Insights into the Conversion of O2 into Two Water Molecules by the Flavoenzyme", Biochemistry 45, 9648-9659 (2006)

Nagpal, A., Valley, M.P., Fitzpatrick, P.F., and Orville, A.M., "Crystal Structures of Nitroalkane Oxidase: Insights into the Reaction Mechanism from a Covalent Complex of the Flavoenzyme Trapped during Turnover" Biochemistry 45, 1138-1150 (2006)

Fitzpatrick, P.F., Orville, A.M., Nagpal, A., & Valley, M.P. "Nitroalkane oxidase, a carbanion-forming flavoprotein homologous to acyl-CoA dehydrogenase" Arch. Biochem. Biophys. 433 157-165 (2005)

Lountos, G.T., Mitchell, K.H., Studts, J.M., Fox B.G. & Orville, A.M., "Crystal structures and functional studies of T4moD, the toluene 4-monooxygenase catalytic effector protein" Biochemistry 44, 7131-7142 (2005)

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