controlled in meso phase crystallization – a method for the structural investigation of membrane proteins控制结晶阶段内消旋-膜蛋白的结构调查的方法.pdf

Controlled In Meso Phase Crystallization – A Method for the Structural Investigation of Membrane Proteins 1 2¤ 2 ¨ 2 ¨ 1 ¨ 2 Jan Kubicek , Ramona Schlesinger , Christian Baeken , Georg Buldt , Frank Schafer , Jorg Labahn * ¨ ¨ 1 QIAGEN GmbH, Hilden, Germany, 2 Molecular Biophysics, Institute of Structural Biology and Biophysics (ISB-2), Research Center Julich, Julich, Germany Abstract We investigated in meso crystallization of membrane proteins to develop a fast screening technology which combines features of the well established classical vapor diffusion experiment with the batch meso phase crystallization, but without premixing of protein and monoolein. It inherits the advantages of both methods, namely (i) the stabilization of membrane proteins in the meso phase, (ii) the control of hydration level and additive concentration by vapor diffusion. The new technology (iii) significantly simplifies in meso crystallization experiments and allows the use of standard liquid handling robots suitable for 96 well formats. CIMP crystallization furthermore allows (iv) direct monitoring of phase transformation ˚ and crystallization events. Bacteriorhodopsin (BR) crystals of high quality and diffraction up to 1.3 A resolution have been obtained in this approach. CIMP and the developed consumables and protocols ha