Nmr As A Structural Tool For Macromolecules

The contemplation of truth and beauty is the proper object for which we were created, which calls forth the most intense desires of the soul, and of which it never tires -Hazlitt In his Nobel lecture Purcell commented that when he saw snow in New England after the discovery of NMR, it appeared like "heaps of protons quietly precessing in earth's magnetic field. " If he were to make the comment in the context of how NMR is being used today, he could have conjured up an image of hydrogen, carbon, and nitrogen nuclei in proteins of an earthbound 8rganism subtly orchestrating a quiet symphony of frequencies, from 150 Hz to 2 kHz, carrying clues to the three-dimensional structure of the macromolecules. The manner in which the basic discoveries of Bloch and Purcell have led to the emergence of NMR, several decades later, as a major technique of biological and medical physics (and chemistry) is a striking example of the power of basic research. It is also a fascinating saga whereby whenever it was felt that the field had reached a plateau, new directions, new technologies, and sometimes serendipity produced new developments that revolutionized the technique and enhanced its capability. As Richard Ernst points out "NMR is intellectually attractive, . . . the practical importance of NMR is enormous, and can justify much of the playful activities of an addicted spectroscopist" (Nobel lecture).

This book presents a critical assessment of progress on the use of nuclear magnetic resonance spectroscopy to determine the structure of proteins, including brief reviews of the history of the field along with coverage of current clinical and in vivo applications. The book, in honor of Oleg Jardetsky, one of the pioneers of the field, is edited by two of the most highly respected investigators using NMR, and features contributions by most of the leading workers in the field. It will be valued as a landmark publication that presents the state-of-the-art perspectives regarding one of today's most important technologies.

The book provides insights into the research of the Kurt Wüthrich laboratories from 1996-2020. During this time period, the technique of nuclear magnetic resonance (NMR) spectroscopy in solution went through several breakthroughs, while maturing into a standard method of structural biology. With the introduction of TROSY (transverse relaxation-optimized spectroscopy), the range of accessible molecular sizes was extended about thirty-fold, and efficient protein structure determination resulted from the demands of the structural genomics initiative. Applications in fundamental biology and biomedicine include studies of prion proteins and prion diseases (TSEs), the SARS-Corona virus proteome, trans-membrane signalling by G protein-coupled receptors (GPCRs), and signal transfer by pheromones.Key publications from the Kurt Wüthrich laboratories are placed in perspective, providing insights into new aspects of NMR spectroscopy in structural biology. In addition to methods development, this includes applications in diverse areas of biological research, such as prion proteins and their role in transmissible spongiform encephalopathies (TSEs), trans-membrane signal transfer by G protein-coupled receptors (GPCRs), structural characterization of the SARS-Corona virus proteome, metabolic-flux profiling in bacterial cultures, and signal transfers by pheromones.

NMR spectroscopy has proven to be a powerful technique to study the structure and dynamics of biological macromolecules. Fundamentals of Protein NMR Spectroscopy is a comprehensive textbook that guides the reader from a basic understanding of the phenomenological properties of magnetic resonance to the application and interpretation of modern multi-dimensional NMR experiments on 15N/13C-labeled proteins. Beginning with elementary quantum mechanics, a set of practical rules is presented and used to describe many commonly employed multi-dimensional, multi-nuclear NMR pulse sequences. A modular analysis of NMR pulse sequence building blocks also provides a basis for understanding and developing novel pulse programs. This text not only covers topics from chemical shift assignment to protein structure refinement, as well as the analysis of protein dynamics and chemical kinetics, but also provides a practical guide to many aspects of modern spectrometer hardware, sample preparation, experimental set-up, and data processing. End of chapter exercises are included to emphasize important concepts. Fundamentals of Protein NMR Spectroscopy not only offer students a systematic, in-depth, understanding of modern NMR spectroscopy and its application to biomolecular systems, but will also be a useful reference for the experienced investigator.

The volume presents a survey of the research by Kurt Wthrich and his associates during the period 1965 to 1994. A selection of reprints of original papers on the use of NMR spectroscopy in structural biology is supplemented with an introduction, which outlines the foundations and the historical development of the use of NMR spectroscopy for the determination of three-dimensional structures of biological macromolecules in solution. The original papers are presented in groups highlighting protein structure determination by NMR, studies of dynamic properties and hydration of biological macromolecules, and practical applications of the NMR methodology in fields such as enzymology, transcriptional regulation, immunosuppression and protein folding.

Provided here are the latest techniques of NMR as applied to the study of proteins, carbohydrates and nucleic acids. The first chapters are devoted to an introduction to NMR and parameters related to molecular structure and molecular interactions. NMR experiments from basic 1D to 2D, 3D and 4D, used in combination with isotopically labelled molecules, are described and a general strategy is presented for biomacromolecular structure determination. Subsequent chapters deal with more advanced principles and techniques and their applications to structural and dynamic processes involving biomacromolecules in solution. Advanced results on peptide, protein, oligosaccharide and nucleic acid structure and recognition are presented.

List of contributors; Nomenclature and abbreviations; Introduction; Sample preparation; Instrumentation and pulse sequences; Resonance assignment strategies for small proteins; Stable isotope labelling and resonance assigments in larger proteins; Effects of chemical exchange on NMR spectra; NMR studies of protein-ligand interactions; NMR of nucleic acids: from spectrum to structure; H NMR studies of oligosaccharides; structure determination from NMR data I. analysis of NMR data; Structure determination from NMR data II. computational approaches; Appendix; Index.

The determination of the three-dimensional structure of a biological molecule is the starting point in the understanding of molecular mechanisms involved in its complex biochemical reactions. The molecular architecture of multimolecular systems such as membranes and chromosomes provides the key to the fascinating field of molecular biology. Stereochemical details of biological macromolecules and their interactions with pharmacological agents form the basis for drug design. Naturally, the study of the structure and function of biological molecules has aroused tremendous interest and investigations in this area are being carried out in a large number of laboratories. The techniques used for this purpose include both experimental methods (X-ray and neutron diffraction measurements, study of NMR, ESR, vibrational and electronic spectra, ORD, CD and dipole moment measurements, biochemical modifications etc. ) and the oretical methods (quantum mechanical and classical potential energy calculations, Monte Carlo simulations and molecular graphics). F or several years now, X-ray diffraction [1] has served as our only source of infor mation on the three-dimensional arrangements of atoms in biopolymers. Fiber-diffrac tion of DNA led to the proposal of the DNA double helix. Fibers of long~hain polymers show ordering in the direction of the fibre-axis but not in the transverse plane. Accurate estimates of the dimensions of helical structures can be made using techniques on the basis of which models of biopolymers can be constructed.

Without computers - no modern NMR; Parametric estimation in 1-D, 2-D, and 3-D NMR; Computational aspects of multinuclear NMR spectroscopy of proteins at NMRFAM; Principles of multidimensional NMR techniques for measurement of J coupling constants; Comparison of the NMR and X-ray structures of hirudin; The application of the linear prediction principle to NMR spectroscopy; NMR data processing and structure calculations using parallel computers; Software approaches for determination of 3-dimensional molecular structures from multi-dimensional NMR; Applicability and limitations of three-dimensional NMR spectroscopy for the study of proteins in solution; The role of selective two-dimensional NMR correlation methods in supplementing computer-supported multiplet analysis by MARCO POLO; Application of maximum entropy methods to NMR spectra of proteins; Pattern recognition in two-dimensional NMR spectra of proteins; The application and development of software tools for the processinf and analysis of heteronuclear multi-dimensional NMR data; Distance geometry in torsion angle space: new developments and applications; Structure determination by NMR: the modeling of NMR parameters as ensemble averages; Time averaged distance restraints in NMR based structural refinement; Analysis of backbone dynamics of interleukin-1 beta; A new version of DADAS (Distance Analysis in Dihedral Angle Space) and its performance; An amateur looks at error analysis in the determination of protein structure by NMR; Structural interpretation of NMR data in the presence of motion; New interactive and automatic algorithms for the assignment of NMR spectra; Outline of a computer program for the analysis of protein NMR spectra; Assignment of the NMR spectra of homologous proteins; Incorporation of internal motion in NMR refinements based on NOESY data; Refinement of three-dimensional protein and DNA structures in solution from NMR data; How to deal with spin-diffusion and internal mobility in biomolecules: a relaxation matrix approach; Interactive computer graphics in the assignment of protein 2D and 3D NMR spectra; Determination of large protein structures from NMR data: definition of the solution structure of the TRP repressor; Interpretation of NMR data in terms of protein structure: summary of a round table discussion; Fast calculation of the relaxation matrix; NMR structures of proteins using stereospecific assignments and relaxation matrix refinement in a hybrid method of distance geometry and simulated annealing; A critique of the interpretation of nuclear Overhauser effects of duplex DNA; Improvement in resolution with nonlinear methods applied to NMR signals from macromolecules; STELLA and CLAIRE: a seraglio of programs for human-aided assignment of 2D 1H NMR spectra of proteins; MolSkop: towards NMR molecular scope; Ribonuclease H: full assignment of backbone proton resonances with heteronuclear 3D NMR and solution structure; Sampling properties of simulated annealing and distance geometry.

This 2nd edition begins with an overview of NMR development and applications in biological systems. It describes recent developments in instrument hardware and methodology. Chapters highlight the scope and limitation of NMR methods. While detailed math and quantum mechanics dealing with NMR theory have been addressed in several well-known NMR volumes, chapter two of this volume illustrates the fundamental principles and concepts of NMR spectroscopy in a more descriptive manner. Topics such as instrument setup, data acquisition, and data processing using a variety of offline software are discussed. Chapters further discuss several routine stategies for preparing samples, especially for macromolecules and complexes. The target market for such a volume includes researchers in the field of biochemistry, chemistry, structural biology and biophysics.