One of the major advancements in NMR spectroscopy of biomolecules has been the development of methods for isotope labeling. At the same time there has been a rapid progress in the number of NMR experiments that utilize such isotope labeled samples. Thus a combination of isotopic labeling and multidimensional, multinuclear experiments has significantly expanded the range of problems in structural biology amenable to NMR.

This book is intended to serve as a compendium of isotope labeling for the biomolecular NMR community providing comprehensive coverage of the existing methods and latest developments along with protocols and practical hints on the various experimental aspects. Researchers in academia or industry will find it useful to have a single book covering a wide range of topics including the emerging areas of large/membrane proteins, RNA, metabolonomics and solid state NMR.

Zusammenfassung

NMR spectroscopy has undergone a revolution in recent years with the advent of several new methods overcoming the problems of sensitivity and resolution. Recent developments in biotechnology have made it easier and economical to introduce 13C, 15N and 2H into proteins and nucleic acids. At the same time, there has been an explosion in the number of NMR experiments that utilize such isotope labeled samples. Thus, a combination of isotopic labeling and multidimensional, multinuclear NMR has opened up new avenues for structural studies of proteins, nucleic acids and their complexes.

This book will focus on recent developments in isotope labeling methods for structural studies of small molecules, peptides, proteins and nucleic acids. The aim of the book is to serve as a compendium of isotope labeling for the biomolecular NMR community providing comprehensive coverage of the existing methods and latest developments along with protocols and practical hints on the various experimental aspects. The book will cover a wide range of topics in isotope labeling under one title including emerging areas of metabolonomics and solid state NMR.

Inhalt

1. Isotope labeling methods for large systems. - 2. Segmental labeling to study multidomain proteins. - 3. Isotope Labeling for Solution and Solid-State NMR Spectroscopy of Membrane Proteins. - 4. Isotope labeling methods for relaxation measurements. - 5. Stereo-array isotope labeling method for studying protein structure and dynamics. - 6. Amino acid selective labeling and unlabeling for protein resonance assignments. - 7. Isotope labeling and segmental labeling of larger RNAs for NMR structural studies. - 8. Isotope Enhanced Approaches in Metabolomics. - 9. Cell-free protein synthesis using E. coli cell extract for NMR studies. - 10. Isotope labeling in insect cells. - 11. Mammalian Expression of Isotopically Labeled Proteins for NMR Spectroscopy.