Protein NMR Techniques
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Protein NMR Techniques

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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.show more

Product details

  • Hardback | 487 pages
  • 157.5 x 231.1 x 33mm | 975.23g
  • Humana Press Inc.
  • Totowa, NJ, United States
  • English
  • Revised
  • 2nd ed. 2004
  • biography
  • 1588292460
  • 9781588292469

Review quote

From reviews of the first edition... "...includes many good practical details which are so often omitted and which workers new to the field have to rediscover..." -Analyst "...an excellent text... a very useful compilation...The limitations and power of the approaches are well presented and should help in experimental design very considerably." -Journal of Medicinal Chemistry "...a collection of excellent reviews that will form a useful contribution to the bookshelf of any NMR lab." -Trends in Biotechnology "...an excellent introduction...Readers will be left with a good understanding of the application of NMR to the study of proteins. " -Quarterly Review of Biologyshow more

Back cover copy

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(TM) 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.show more

Table of contents

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. O'Leary 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 Indexshow more

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