Protein Crystallization Strategies for Structural Genomics

During the 1980s and early 1990s, we observed a tremendous breakthrough in our understanding of the physical chemistry of macromolecular crystallization due to the pioneering efforts of several leaders in the field. However, the determination of protein 3-dimensional structure was still a slow process, often taking years to complete. In the mid to late 1990s, a new renaissance of protein crystallography occurred with the development of increased throughput nanovolume crystallization robotic methods, microfluidic counter diffusion, creative screens and optimization techniques, crystal imaging, and novel crystallization methods for the very challenging membrane proteins and large macromolecular complexes. In the fast paced world of high throughput protein crystallization, this work was established by many of the authors in this book and the areas are comprehensively covered. Protein Crystallization Strategies for Structural Genomics will be highly valuable to those experimentalist interested in understanding, reproducing, or expanding upon the recent innovations. It will be an essential reference for the many small and medium size laboratories that are starting or are planning to adopt these systems and approaches. The clarity, thoroughness, and simplicity of the chapters are impressive. The best analogy for this compendium is the early days of gene sequencing when technology development was critical in moving radioactive gel sequencing to fluorescent capillary automated methods. This book is the beginning of a similar movement, initiating the vision of rapid macromolecular structure determination for providing a deeper understanding of the molecular science of life. What this book accomplishes and that many other similar books miss are the critical details necessary to understand, and further expand upon protein crystallization strategies for structural biology. -- --Raymond C. Stevens, Professor, The Scripps Research Institute, La Jolla, CA

From Crystallography Reviews-- Protein Crystallization Strategies for Structural Genomics comprises nine main chapters, each corresponding to different aspects or methods of protein crystallization. It spans a general introduction to the main problems of automation of this process exploring some technicalities of the equipment and methods and also encompasses topics such as image processing and recognition. The short introductory chapter by C.G. Edmonds and J.C. Norwell outlines the origins of the USA National Institute of General Medical Sciences (NIGMS) Protein Structure Initiative (PSI): its philosophy, the different phases of development and any bottlenecks so far encountered. The book really starts with a very substantial second chapter: A Guide to Automation and Data Handling in Protein Crystallization by Bernhard Rupp. This is an excellent and independent introduction to most of the problems of automation and high-throughput (HT) in protein crystallization. The real strength of this chapter is its persistently broad context to the subject. B. Rupp has embarked here on a very ambitious presentation of practically all aspects of protein crystallization, evaluating at the same time applicability of automation-HT in both the genomics and the academic research environments. The conflict between broad HT and low throughput evaluation perspectives makes this chapter very helpful to different types of readers with diverse research interests and aims. The first table of this chapter lists the key environmental factors in academia in contrast to the industry-like research efforts and their impact on deployment of automation. Although it is of rather general character, it provides useful checkpoints for the non-genomics laboratory in considering whether to embark on the automation of crystallization and which of its stages could be most suitable for implementation. There is also an early and honest warning about a lack of an immediate guarantee of automation-HT for high structural output: Figure 2.1 illustrates success statistics for PSI and indicates that its cumulative success rate from targets-to-genes is about 13% for the best centres while on average it lingers at around ca 0.7% (although it may be higher now! these data relate to the first quarter of 2004). Considering this relatively low success rate and undeniably high costs of HT instrumentation, B. Rupp is trying to strip off most of the HT-related myths to provide regular laboratories with a very honest and useful process of evaluation which and when HT-robots could be implemented to provide real gains for the research output. The automation of particular stages of protein crystallization should be carefully tailored to the needs and specificity of a particular laboratory. B. Rupp warns that an over-enthusiastic automation of one particular crystallization step without parallel measures can easily lead to upstream or downstream bottlenecks that will tamper with the overall progress: ...high-throughput crystallization of even a few proteins per day is not sustainable without careful planning of the entire automation setup and thorough operations review. -- The full review is published in Crystallography Reviews, Vol. 15, No. 2, April-June 2009, 135-139 --Andrzej Marek Brzozowski, University of York, UK