结构生物学sturcture biology explore-7.pdfVIP

review 740 nature structural biology ? volume 7 number 9 ? september 2000 Proteins are dynamic molecules that often undergo conforma- tional changes while performing their specific functions, such as an enzyme reaction or ligand binding. The dynamic properties intrinsic to a protein structure may provide information on the location and the energetics of the conformational change process, and are thus the focus of many biophysical studies. Nuclear mag- netic resonance (NMR) spectroscopy can be used to monitor the dynamic behavior of a protein at a multitude of specific sites. Moreover, protein movements on a broad range of timescales can be monitored using various types of NMR experiments — nuclear spin relaxation rate measurements report the internal motions on fast (subnanoseconds) and slow (microseconds to milliseconds) timescales as well as the overall rotational diffusion of the molecule (5–50 nanoseconds), whereas rates of magnetiza- tion transfer among protons with different chemical shifts and proton exchange report movements of protein domains on the very slow timescales (milliseconds to days). These features make NMR a unique and powerful tool in studying protein dynamics related to protein functions, and there has been a tremendous growth in these applications since the review by Lewis Kay1 in Nature Structural Biology in 1998. In parallel with new applica- tions, there have been important developments in experimental methods that significantly increased the accuracy of the informa- tion obtained, and thus expanded the range of questions that can be addressed. In the first section, we will focus in detail on the methods used to analyze conformational changes with NMR spectroscopy. The remainder of the review will provide examples of the usefulness of this technology. Methodological advances Fast protein dynamics. Information about site-specific internal dynamics on the subnanosecond timescale derives primarily from model-free analyses2,3 (Fig. 1). From suc