X-ray crystallography marries spectroscopy to unveil structure and function of biological macromolecules.

摘要

X-ray crystallography of biological macromolecules has provided a wealth of information that has shaped and advanced many fields, including biology, biochemistry, medicine and medicinal chemistry since the crystal structure of sperm whale myoglobin was solved by Kendrew in 1957 [1]. There are now almost 62,000 X-ray structures deposited in the Protein Databank (as of February 2011) and, given the upwards trend (Fig. 1), many more structures will be determined in the coming years. The remarkable success in macromolecular structure determination since the mid-1990s is due to five major factors: i) the ability to generate sufficient pure protein for crystallization using heterologous expression systems and easy purification via affinity chromatography, ii) the development and availability of automation at every stage of the structure determination process, from protein production to crystallization and diffraction data collection, iii) the accessibility of intense X-ray beams at 2nd and 3rd generation synchrotron sources and, more recently, the development of high intensity microfocus beams enabling the determination of structures from crystals only a few microns in size, iv) the development of cryo-methods that increase the lifetime of the crystal within the X-ray beam, and, finally, v) the expanding knowledge of cellular processes and protein interactions via proteomics and interactomics that has led to the identification of new, interesting proteins and protein complexes for structural studies.