X-ray Transparent Microfluidic Chip for Mesophase-Based Crystallization of Membrane Proteins and On-Chip Structure Determination

Khvostichenko, Daria S.; Schieferstein, Jeremy M.; Pawate, Ashtamurthy S.; Laible, Philip D.; Kenis, Paul J.  A.

doi:10.1021/cg5011488

X-ray Transparent Microfluidic Chip for Mesophase-Based Crystallization of Membrane Proteins and On-Chip Structure Determination

Journal Article · Wed Oct 01 00:00:00 EDT 2014 · Crystal Growth and Design

DOI:https://doi.org/10.1021/cg5011488· OSTI ID:1393969

Khvostichenko, Daria S.; Schieferstein, Jeremy M.; Pawate, Ashtamurthy S.; Laible, Philip D.; Kenis, Paul J. A.

Crystallization from lipidic mesophase matrices is a promising route to diffraction-quality crystals and structures of membrane proteins. The microfluidic approach reported here eliminates two bottlenecks of the standard mesophase-based crystallization protocols: (i) manual preparation of viscous mesophases and (ii) manual harvesting of often small and fragile protein crystals. In the approach reported here, protein-loaded mesophases are formulated in an X-ray transparent microfluidic chip using only 60 nL of the protein solution per crystallization trial. The X-ray transparency of the chip enables diffraction data collection from multiple crystals residing in microfluidic wells, eliminating the normally required manual harvesting and mounting of individual crystals. We validated our approach by on-chip crystallization of photosynthetic reaction center, a membrane protein from Rhodobacter sphaeroides, followed by solving its structure to a resolution of 2.5 Å using X-ray diffraction data collected on-chip under ambient conditions. A moderate conformational change in hydrophilic chains of the protein was observed when comparing the on-chip, room temperature structure with known structures for which data were acquired under cryogenic conditions.

Research Organization:: Argonne National Laboratory (ANL)

Sponsoring Organization:: USDOE Advanced Research Projects Agency - Energy (ARPA-E)

DOE Contract Number:: AC02-06CH11357

OSTI ID:: 1393969

Journal Information:: Crystal Growth and Design, Journal Name: Crystal Growth and Design Journal Issue: 10 Vol. 14; ISSN 1528-7483

Publisher:: American Chemical Society

Country of Publication:: United States

Language:: English

References (44)

Radiation damage in macromolecular crystallography: what is it and why should we care? Garman, Elspeth F. Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 4 https://doi.org/10.1107/S0907444910008656	journal	March 2010
A plug-based microfluidic system for dispensing lipidic cubic phase (LCP) material validated by crystallizing membrane proteins in lipidic mesophases Li, Liang; Fu, Qiang; Kors, Christopher A. Microfluidics and Nanofluidics, Vol. 8, Issue 6 https://doi.org/10.1007/s10404-009-0512-8	journal	October 2009
In situ X-ray analysis of protein crystals in low-birefringent and X-ray transmissive plastic microchannels Ng, Joseph D.; Clark, Peter J.; Stevens, Raymond C. Acta Crystallographica Section D Biological Crystallography, Vol. 64, Issue 2 https://doi.org/10.1107/S0907444907060064	journal	January 2008
Lipidic cubic phase technologies for membrane protein structural studies Cherezov, Vadim Current Opinion in Structural Biology, Vol. 21, Issue 4 https://doi.org/10.1016/j.sbi.2011.06.007	journal	August 2011
Diversity and modularity of G protein-coupled receptor structures Katritch, Vsevolod; Cherezov, Vadim; Stevens, Raymond C. Trends in Pharmacological Sciences, Vol. 33, Issue 1 https://doi.org/10.1016/j.tips.2011.09.003	journal	January 2012
Recent Progress in the Structure Determination of GPCRs, a Membrane Protein Family with High Potential as Pharmaceutical Targets Cherezov, Vadim; Abola, Enrique; Stevens, Raymond C. Methods in Molecular Biology https://doi.org/10.1007/978-1-60761-762-4_8	book	January 2010
Laue Crystallography for Studying Rapid Reactions Ren, Z.; Moffat, K. Journal of Synchrotron Radiation, Vol. 1, Issue 1 https://doi.org/10.1107/s0909049594006692	journal	October 1994
From 3D cell culture to organs-on-chips Huh, Dongeun; Hamilton, Geraldine A.; Ingber, Donald E. Trends in Cell Biology, Vol. 21, Issue 12 https://doi.org/10.1016/j.tcb.2011.09.005	journal	December 2011
A microfluidic approach for protein structure determination at room temperature via on-chip anomalous diffraction Perry, Sarah L.; Guha, Sudipto; Pawate, Ashtamurthy S. Lab on a Chip, Vol. 13, Issue 16 https://doi.org/10.1039/c3lc50276g	journal	January 2013
Lipidic Sponge Phase Crystallization of Membrane Proteins Wadsten, Pia; Wöhri, Annemarie B.; Snijder, Arjan Journal of Molecular Biology, Vol. 364, Issue 1 https://doi.org/10.1016/j.jmb.2006.06.043	journal	November 2006
From screen to structure with a harvestable microfluidic device Stojanoff, Vivian; Jakoncic, Jean; Oren, Deena A. Acta Crystallographica Section F Structural Biology and Crystallization Communications, Vol. 67, Issue 8 https://doi.org/10.1107/S1744309111024456	journal	July 2011
Room to Move: Crystallizing Membrane Proteins in Swollen Lipidic Mesophases Cherezov, Vadim; Clogston, Jeffrey; Papiz, Miroslav Z. Journal of Molecular Biology, Vol. 357, Issue 5 https://doi.org/10.1016/j.jmb.2006.01.049	journal	April 2006
Integrated microfluidic reactors Lin, Wei-Yu; Wang, Yanju; Wang, Shutao Nano Today, Vol. 4, Issue 6 https://doi.org/10.1016/j.nantod.2009.10.007	journal	December 2009
Behavior of capillary valves in centrifugal microfluidic devices prepared by three-dimensional printing Moore, Jessica L.; McCuiston, Austin; Mittendorf, Isaac Microfluidics and Nanofluidics, Vol. 10, Issue 4 https://doi.org/10.1007/s10404-010-0721-1	journal	November 2010
Chaotic Mixer for Microchannels Stroock, A. D. Science, Vol. 295, Issue 5555 https://doi.org/10.1126/science.1066238	journal	January 2002
Toward on-chip X-ray analysis Greaves, Eduardo D.; Manz, Andreas Lab on a Chip, Vol. 5, Issue 4 https://doi.org/10.1039/b415836a	journal	January 2005
Pros and cons of cryocrystallography: should we also collect a room-temperature data set? Dunlop, Kirsty V.; Irvin, Randall T.; Hazes, Bart Acta Crystallographica Section D Biological Crystallography, Vol. 61, Issue 1 https://doi.org/10.1107/S0907444904027179	journal	December 2004
Microfluidics for drug discovery and development: From target selection to product lifecycle management Kang, L. Drug Discovery Today, Vol. 13, Issue 1-2 https://doi.org/10.1016/j.drudis.2007.10.003	journal	January 2008
Serial Femtosecond Crystallography of G Protein-Coupled Receptors Liu, W.; Wacker, D.; Gati, C. Science, Vol. 342, Issue 6165 https://doi.org/10.1126/science.1244142	journal	December 2013
ChipX: A Novel Microfluidic Chip for Counter-Diffusion Crystallization of Biomolecules and in Situ Crystal Analysis at Room Temperature Pinker, Franziska; Brun, Mathieu; Morin, Pierre Crystal Growth & Design, Vol. 13, Issue 8 https://doi.org/10.1021/cg301757g	journal	July 2013
Accessing protein conformational ensembles using room-temperature X-ray crystallography Fraser, J. S.; van den Bedem, H.; Samelson, A. J. Proceedings of the National Academy of Sciences, Vol. 108, Issue 39 https://doi.org/10.1073/pnas.1111325108	journal	September 2011
Lipidic Cubic Phase Crystal Structure of the Photosynthetic Reaction Centre from Rhodobacter sphaeroides at 2.35Å Resolution Katona, Gergely; Andréasson, Ulf; Landau, Ehud M. Journal of Molecular Biology, Vol. 331, Issue 3 https://doi.org/10.1016/S0022-2836(03)00751-4	journal	August 2003
X-CHIP: an integrated platform for high-throughput protein crystallization and on-the-chip X-ray diffraction data collection Kisselman, Gera; Qiu, Wei; Romanov, Vladimir Acta Crystallographica Section D Biological Crystallography, Vol. 67, Issue 6 https://doi.org/10.1107/S0907444911011589	journal	May 2011
The plug-based nanovolume Microcapillary Protein Crystallization System (MPCS) Gerdts, Cory J.; Elliott, Mark; Lovell, Scott Acta Crystallographica Section D Biological Crystallography, Vol. 64, Issue 11 https://doi.org/10.1107/S0907444908028060	journal	October 2008
A robotic system for crystallizing membrane and soluble proteins in lipidic mesophases Cherezov, Vadim; Peddi, Avinash; Muthusubramaniam, Lalitha Acta Crystallographica Section D Biological Crystallography, Vol. 60, Issue 10 https://doi.org/10.1107/S0907444904019109	journal	September 2004
X-ray microfocussing combined with microfluidics for on-chip X-ray scattering measurements Barrett, Ray; Faucon, Marc; Lopez, John Lab on a Chip, Vol. 6, Issue 4 https://doi.org/10.1039/b517055a	journal	January 2006
From Macrofluidics to Microfluidics for the Crystallization of Biological Macromolecules ^† Sauter, Claude; Dhouib, Kaouthar; Lorber, Bernard Crystal Growth & Design, Vol. 7, Issue 11 https://doi.org/10.1021/cg700955f	journal	November 2007
Crystallizing membrane proteins using lipidic mesophases Caffrey, Martin; Cherezov, Vadim Nature Protocols, Vol. 4, Issue 5 https://doi.org/10.1038/nprot.2009.31	journal	April 2009
Chapter 4 Monoacylglycerols Caffrey, M.; Lyons, J.; Smyth, T. Current Topics in Membranes https://doi.org/10.1016/S1063-5823(09)63004-0	book	January 2009
How many drug targets are there? Overington, John P.; Al-Lazikani, Bissan; Hopkins, Andrew L. Nature Reviews Drug Discovery, Vol. 5, Issue 12 https://doi.org/10.1038/nrd2199	journal	December 2006
Controlled In Meso Phase Crystallization – A Method for the Structural Investigation of Membrane Proteins Kubicek, Jan; Schlesinger, Ramona; Baeken, Christian PLoS ONE, Vol. 7, Issue 4 https://doi.org/10.1371/journal.pone.0035458	journal	April 2012
Lipidic cubic phase injector facilitates membrane protein serial femtosecond crystallography Weierstall, Uwe; James, Daniel; Wang, Chong Nature Communications, Vol. 5, Issue 1 https://doi.org/10.1038/ncomms4309	journal	February 2014
Lipidic Cubic Phase Crystal Structure of the Photosynthetic Reaction Centre from Rhodobacter sphaeroides at 2.35Å Resolution Katona, Gergely; Andréasson, Ulf; Landau, Ehud M. Journal of Molecular Biology, Vol. 331, Issue 3 https://doi.org/10.1016/s0022-2836(03)00751-4	journal	August 2003
A microfluidic processor for gene expression profiling of single human embryonic stem cells Zhong, Jiang F.; Chen, Yan; Marcus, Joshua S. Lab Chip, Vol. 8, Issue 1 https://doi.org/10.1039/B712116D	journal	January 2008
Microfluidic chips for the crystallization of biomacromolecules by counter-diffusion and on-chip crystal X-ray analysis Dhouib, Kaouthar; Khan Malek, Chantal; Pfleging, Wilhelm Lab on a Chip, Vol. 9, Issue 10 https://doi.org/10.1039/b819362b	journal	January 2009
Monoolein Lipid Phases as Incorporation and Enrichment Materials for Membrane Protein Crystallization Wallace, Ellen; Dranow, David; Laible, Philip D. PLoS ONE, Vol. 6, Issue 8 https://doi.org/10.1371/journal.pone.0024488	journal	August 2011
How root-mean-square distance (r.m.s.d.) values depend on the resolution of protein structures that are compared Carugo, Oliviero Journal of Applied Crystallography, Vol. 36, Issue 1 https://doi.org/10.1107/S0021889802020502	journal	January 2003
Shear Rheology of Lyotropic Liquid Crystals: A Case Study Mezzenga, Raffaele; Meyer, Cedric; Servais, Colin Langmuir, Vol. 21, Issue 8 https://doi.org/10.1021/la046964b	journal	April 2005
Laue Crystallography for Studying Rapid Reactions Ren, Z.; Moffat, K. Journal of Synchrotron Radiation, Vol. 1, Issue 1 https://doi.org/10.1107/S0909049594006692	journal	October 1994
Radiation damage in macromolecular crystallography: what is it and why should we care? Garman, Elspeth F. Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 4 https://doi.org/10.1107/s0907444910008656	journal	March 2010
Fabrication of X-ray compatible microfluidic platforms for protein crystallization Guha, Sudipto; Perry, Sarah L.; Pawate, Ashtamurthy S. Sensors and Actuators B: Chemical, Vol. 174 https://doi.org/10.1016/j.snb.2012.08.048	journal	November 2012
Two-layer multiplexed peristaltic pumps for high-density integrated microfluidics Cole, Matthew C.; Desai, Amit V.; Kenis, Paul J. A. Sensors and Actuators B: Chemical, Vol. 151, Issue 2, p. 384-393 https://doi.org/10.1016/j.snb.2010.07.012	journal	January 2011
Nonlinear Optical Imaging of Integral Membrane Protein Crystals in Lipidic Mesophases Kissick, David J.; Gualtieri, Ellen J.; Simpson, Garth J. Analytical Chemistry, Vol. 82, Issue 2 https://doi.org/10.1021/ac902139w	journal	January 2010
From The Cover: Systematic investigation of protein phase behavior with a microfluidic formulator Hansen, C. L.; Sommer, M. O. A.; Quake, S. R. Proceedings of the National Academy of Sciences, Vol. 101, Issue 40, p. 14431-14436 https://doi.org/10.1073/pnas.0405847101	journal	September 2004

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