When I was asked to edit the second edition of Protein NMR Techniques, my first thought was that the time was ripe for a new edition. The past several years have seen a surge in the development of novel methods that are truly revolutionizing our ability to characterize biological macromolecules in terms of speed, accuracy, and size limitations. I was particularly excited at the prospect of making these techniques accessible to all NMR labs and for the opportunity to ask the experts to divulge their hints and tips and to write, practically, about the methods. I commissioned 19 chapters with wide scope for Protein NMR Techniques, and the volume has been organized with numerous themes in mind. Chapters 1 and 2 deal with recombinant protein expression using two organisms, E. coli and P. pastoris, that can produce high yields of isotopically labeled protein at a reasonable cost. Staying with the idea of isotopic labeling, Chapter 3 describes methods for perdeuteration and site-specific protonation and is the first of several chapters in the book that is relevant to studies of higher molecular weight systems.
A different, but equally powerful, method that uses molecular biology to "edit" the spectrum of a large molecule using segmental labeling is presented in Chapter 4. Having successfully produced a high molecular weight target for study, the next logical step is data acquisition. Hence, the final chapter on this theme, Chapter 5, describes TROSY methods for stru- ural studies.
Screening and Optimizing Protein Production in E. coli Lorraine Hewitt and James M. McDonnell Isotopic Labeling of Recombinant Proteins From the Methylotrophic Yeast Pichia pastoris Andrew R. Pickford and Joanne M. OLeary Perdeuteration/Site-Specific Protonation Approaches for High-Molecular-Weight Proteins Stephen Matthews Segmental Isotopic Labeling for Structural Biological Applications of NMR David Cowburn, Alexander Shekhtman, Rong Xu, Jennifer J. Ottesen, and Tom W. Muir TROSY-Based Correlation and NOE Spectroscopy for NMR Structural Studies of Large Proteins Guang Zhu, Youlin Xia, Donghai Lin, and Xiaolian Gao Media for Studies of Partially Aligned States Kieran Fleming and Stephen Matthews Residual Dipolar Couplings in Protein Structure Determination Eva de Alba and Nico Tjandra Projection Angle Restraints for Studying Structure and Dynamics of Biomolecules Christian Griesinger, Wolfgang Peti, Jens Meiler, and Raphael Bruschweiler Characterizing Domain Interfaces by NMR Luke M. Rooney, Sachchidanand, and Jorn M. Werner Characterization of the Overall Rotational Diffusion of a Protein From 15N Relaxation Measurements and Hydrodynamic Calculations Jennifer Blake Hall, Olivier Walker, and David Fushman TROSY-Based NMR Experiments for the Study of Macromolecular Dynamics and Hydrogen Bonding Guang Zhu, Youlin Xia, Donghai Lin, and Xiaolian Gao Measurement of Intermediate Exchange Phenomena James G. Kempf and J. Patrick Loria NMR Studies of Partially Folded Molten-Globule States Christina Redfield Structure Determination of Protein Complexes by NMR Daniel Nietlispach, Helen R. Mott, Katherine M. Stott, Peter R. Nielsen, Abarna Thiru, and Ernest D. Laue NMR Studies of Protein-Nucleic Acid Interactions Gabriele Varani, Yu Chen, andThomas C. Leeper Using NMRView to Visualize and Analyze the NMR Spectra of Macromolecules Bruce A. Johnson Automated NMR Structure Calculation With CYANA Peter Guntert NOE Assignment With ARIA 2.0: The Nuts and Bolts Michael Habeck, Wolfgang Rieping, Jens P. Linge, and Michael Nilges Membrane Protein Structure Determination Using Solid-State NMR Anthony Watts, Suzana K. Straus, Stephan L. Grage, Miya Kamihira, Yuen Han Lam, and Xin Zhao Index