Optimization of Protein Crystallization: The OptiCryst Project
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- 作者：Alfonso Garcia-Caballero ; Jose A. Gavira ; Estela Pineda-Molina ; Naomi E. Chayen ; Lata Govada ; Sahir Khurshid ; Emmanuel Saridakis ; Attia Boudjemline ; Marcus J. Swann ; Patrick Shaw Stewart ; Richard A. Briggs ; Stefan A. Kolek ; Dominik Oberthuer ; Karsten Dierks ; Christian Betzel ; Martha Santana ; Jeanette R. Hobbs ; Paul Thaw ; Tony J. Savill ; Jeroen R. Mesters ; Rolf Hilgenfeld ; Nicklas Bonander ; Roslyn M. Bill
- 刊名：Crystal Growth & Design
- 出版年：2011
- 出版时间：June 1, 2011
- 年：2011
- 卷：11
- 期：6
- 页码：2112-2121
- 全文大小：1054K
- 年卷期：v.11,no.6(June 1, 2011)
- ISSN：1528-7505
文摘
Protein crystallization has gained a new strategic and commercial relevance in the postgenomic era due to its pivotal role in structural genomics. Producing high quality crystals has always been a bottleneck to efficient structure determination, and this problem is becoming increasingly acute. This is especially true for challenging, therapeutically important proteins that typically do not form suitable crystals. The OptiCryst consortium has focused on relieving this bottleneck by making a concerted effort to improve the crystallization techniques usually employed, designing new crystallization tools, and applying such developments to the optimization of target protein crystals. In particular, the focus has been on the novel application of dual polarization interferometry (DPI) to detect suitable nucleation; the application of in situ dynamic light scattering (DLS) to monitor and analyze the process of crystallization; the use of UV-fluorescence to differentiate protein crystals from salt; the design of novel nucleants and seeding technologies; and the development of kits for capillary counterdiffusion and crystal growth in gels. The consortium collectively handled 60 new target proteins that had not been crystallized previously. From these, we generated 39 crystals with improved diffraction properties. Fourteen of these 39 were only obtainable using OptiCryst methods. For the remaining 25, OptiCryst methods were used in combination with standard crystallization techniques. Eighteen structures have already been solved (30% success rate), with several more in the pipeline.