Screening and Optimizing Protein Production in E. coli -- Isotopic Labeling of Recombinant Proteins from the Methylotrophic Yeast Pichia pastoris -- Perdeuteration/Site-Specific Protonation Approaches for High-Molecular-Weight Proteins -- Segmental Isotopic Labeling for Structural Biological Applications of NMR -- TROSY-Based Correlation and NOE Spectroscopy for NMR Structural Studies of Large Proteins -- Media for Studies of Partially Aligned States -- Residual Dipolar Couplings in Protein Structure Determination -- Projection Angle Restraints for Studying Structure and Dynamics of Biomolecules -- Characterizing Domain Interfaces by NMR -- Characterization of the Overall Rotational Diffusion of a Protein From 15N Relaxation Measurements and Hydrodynamic Calculations -- TROSY-Based NMR Experiments for the Study of Macromolecular Dynamics and Hydrogen Bonding -- Measurement of Intermediate Exchange Phenomena -- NMR Studies of Partially Folded Molten-Globule States -- Structure Determination of Protein Complexes by NMR -- NMR Studies of Protein-Nucleic Acid Interactions -- Using NMRView to Visualize and Analyze the NMR Spectra of Macromolecules -- Automated NMR Structure Calculation With CYANA -- NOE Assignment With ARIA 2.0 -- Membrane Protein Structure Determination Using Solid-State NMR.

Carrying on in the tradition of its much praised earlier edition, A. Kristina Downing has selected new protocols describing the many recent advances in NMR methodology that are revolutionizing the field. Described in step-by-step detail by established hands-on experts, these readily reproducible techniques include methods for high-level recombinant protein expression using sophisticated isotopic labeling strategies, TROSY methods for the study of structure and dynamics, and methods for the acquisition and interpretation of residual dipolar coupling data. There are also applications of dynamics measurements on multiple timescales, new developments in NMR data analysis, structure calculation protocols, and a comprehensive chapter on solid-state methods for the study of membrane proteins. The protocols follow the successful Methods in Molecular Biology™ series format, each offering detailed laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. State-of-the-art and highly practical, Protein NMR Techniques, 2nd Ed., makes accessible to all NMR laboratories proven techniques that are powerfully transforming the speed, accuracy, and size limit of our ability to characterize biological macromolecules.