Serial Millisecond Crystallography of Membrane Proteins
Serial femtosecond crystallography (SFX) at X-ray free-electron lasers (XFELs) is a powerful method to determine high-resolution structures of pharmaceutically relevant membrane proteins. Recently, the technology has been adapted to carry out serial millisecond crystallography (SMX) at synchrotron sources, where beamtime is more abundant. In an injector-based approach, crystals grown in lipidic cubic phase (LCP) or embedded in viscous medium are delivered directly into the unattenuated beam of a microfocus beamline. Pilot experiments show the application of microjet-based SMX for solving the structure of a membrane protein and compatibility of the method with de novo phasing. Planned synchrotron upgrades, faster detectors and software developments will go hand-in-hand with developments at free-electron lasers to provide a powerful methodology for solving structures from microcrystals at room temperature, ligand screening or crystal optimization for time-resolved studies with minimal or no radiation damage.
KeywordsSerial crystallography High viscosity injector LCP injector Synchrotron Membrane proteins
The work was financially supported by A Co-fund PSI Fellowship (to P.N.) and the SNSF project grant 31003A_141235 and 31003A_159558 (toJ.S.).
- Demirci H, Sierra RG, Laksmono H, Shoeman RL, Botha S, Barends TRM, Nass K, Schlichting I, Doak RB, Gati C, Williams GJ, Boutet S, Messerschmidt M, Jogl G, Dahlberg AE, Gregory ST, Bogan MJ (2013) Serial femtosecond X-ray diffraction of 30S ribosomal subunit microcrystals in liquid suspension at ambient temperature using an X-ray free-electron laser. Acta Crystallogr Sect F Struct Biol Cryst Commun 69(9):1066–1069CrossRefPubMedPubMedCentralGoogle Scholar
- Johnson I, Bergamaschia A, Billich H, Cartier S, Dinapoli R, Greiffenberg D, Guizar-Sicairos M, Henrich B, Jungmann J, Mezza D, Mozzanica A, Schmitt B, Shi X, Tinti G (2014) Eiger: a single-photon counting x784 ray detector. J Instrum 9Google Scholar