2D femtosecond crystallography: X-ray free electron laser studies on two-dimensional crystals of membrane proteins

 
Schertler    Prof. Dr.
Gebhard F. X. Schertler
Paul Scherrer Institut
OFLC/109
CH-5232 Villigen PSI
Laboratory of Biomolecular Research, PSI
+41 56 310 4265 
Project starts   1.3.2012
Project ends    1.3.2015
     
Goals   We are focusing on G protein coupled receptors (GPCR) that are the key players in cell signaling process. We are interested in structures of GPCRs in complex with different signaling partners to understand the basis of signal transduction.  Currently we are in the process to obtain 2D crystals of the light-sensitive rhodopsin in complex with G protein. Once we find the 2D crystallization conditions, we will optimize the 2D crystals and use the XFEL technique to image these 2D crystals. Our major goals are (1) to establish a robust and reproducible pipeline to obtain well-diffracting 2D crystals of GPCR complexes, and (2) to develop a straight-forward procedure for data processing and structure determination by X-ray free-electron laser.
     
Contact  
Ching-Ju Tsai
Paul Scherrer Institut
OFLG/301
5232 VIlligen-PSI
+4156 310 4032
Xiao-Dan Li
Paul Scherrer Institut
OFLC/104
5232 VIlligen-PSI
+4156 310 4469
Gebhard Schertler
Paul Scherrer Institut
OFLC/109
CH-5232 Villigen PSI
+41 56 310 4265  
 
     
Abstract    Our research focuses on understanding the structure-function relationship of membrane proteins. 2D crystals of membrane proteins obtained from the cell membrane-like environment most likely to maintain their functional and structural integrity and they are suitable for structural determination classically using cryo-electron microscopy. The fast development of X-ray free electron laser (XFEL) imaging technique provides a new opportunity for us using 2D membrane protein crystals to obtain structural information. XFEL not only captures the structural information at femtosecond time-scale before radiation damage can occur. We also will obtain direct phase information by using the coherent property of the FEL X-ray pulse. The structural information of 2D membrane protein crystals also contains the dynamic properties of the studied protein in contrast to conventional cryo-electron microscopy and X-ray crystallography of frozen-hydrated specimens. 
     
Publications   Venkatakrishnan, A.J., Deupi, X., Lebon, G., Tate, C.G., Schertler, G.F., and Babu, M.M. (2013) Molecular signatures of G-protein-coupled receptors. Nature 494, 185-194 (10.1038/nature11896) Venkatakrishnan-2013 (2.16 MB).
Panneels, V., et al., Time-resolved structural studies with serial crystallography: A new light on retinal proteins. Struct. Dynam., 2015. 2(4), DOI: 10.1063/1.4922774
Wu, W., et al., Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser. Acta Crystallogr. Sect. F Struct. Biol. Cryst. Commun., 2015. 71: p. 856-860, DOI: 10.1107/s2053230x15009966
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