Protein NMR Spectroscopy

Nuclear Magnetic Resonance (NMR) spectroscopy, a physical phenomenon based upon the magnetic properties of certain atomic nuclei, has found a wide range of applications in life sciences over recent decades. This up-to-date volume covers NMR techniques and their application to proteins, with a focus on practical details. Providing newcomers to NMR with practical guidance to carry out successful experiments with proteins and analyze the resulting spectra, those familiar with the chemical applications of NMR will also find it useful in understanding the special requirements of protein NMR.

Nuclear Magnetic Resonance (NMR) spectroscopy, a physical phenomenon based upon the magnetic properties of certain atomic nuclei, has found a wide range of applications in life sciences over recent decades. This up-to-date volume covers NMR techniques and their application to proteins, with a focus on practical details. Providing newcomers to NMR with practical guidance to carry out successful experiments with proteins and analyze the resulting spectra, those familiar with the chemical applications of NMR will also find it useful in understanding the special requirements of protein NMR.

List of Contributors xiiiIntroduction 1Lu-Yun Lian and Gordon RobertsReferences 41 Sample Preparation, Data Collection and Processing5Frederick W. Muskett1.1 Introduction 51.2 Sample Preparation 51.3 Data Collection 111.4 Data Processing 172 Isotope Labelling 23Mitsuhiro Takeda and Masatsune Kainosho2.1 Introduction 232.2 Production Methods for Isotopically Labelled Proteins 242.3 Uniform Isotope Labelling of Proteins 292.4 Selective Isotope Labelling of Proteins 322.5 Segmental Labelling 372.6 SAIL Methods 382.7 Concluding Remarks 453 Resonance Assignments 55Lu-Yun Lian and Igor L. Barsukov3.1 Introduction 553.2 Resonance Assignment of Unlabelled Proteins 563.3 15N-Edited Experiments 603.4 Triple Resonance 623.5 Side-Chain Assignments 774 Measurement of Structural Restraints 83Geerten W. Vuister, Nico Tjandra, Yang Shen, Alex Grishaev andStephan Grzesiek4.1 Introduction 834.2 NOE-Based Distance Restraints 844.3 Dihedral Restraints Derived from J-Couplings 964.4 Hydrogen Bond Restraints 1034.5 Orientational Restraints 1074.6 Chemical Shift Structural Restraints 1294.7 Solution Scattering Restraints 1375 Calculation of Structures from NMR Restraints 159Peter Guntert5.1 Introduction 1595.2 Historical Development 1615.3 Structure Calculation Algorithms 1645.4 Automated NOE Assignment 1735.5 Nonclassical Approaches 1785.6 Fully Automated Structure Analysis 1816 Paramagnetic Tools in Protein NMR 193Peter H.J. Keizers and Marcellus Ubbink6.1 Introduction 1936.2 Types of Restraints 1946.3 What Metals to Use? 2006.4 Paramagnetic Probes 2036.5 Examples 2096.6 Conclusions and Perspective 2127 Structural and Dynamic Information on Ligand Binding221Gordon Roberts7.1 Introduction 2217.2 Fundamentals of Exchange Effects on NMR Spectra 2227.3 Measurement of Equilibrium and Rate Constants 2297.4 Detecting Binding - NMR Screening 2387.5 Mechanistic Information 2417.6 Structural Information 2468 Macromolecular Complexes 269Paul C. Driscoll8.1 Introduction 2698.2 Spectral Simplification through Differential IsotopeLabelling 2708.3 Basic NMR Characterisation of Complexes 2738.4 3D Structure Determination of MacromolecularProtein-Ligand Complexes 2778.5 Literature Examples 2979 Studying Partially Folded and Intrinsically DisorderedProteins Using NMR Residual Dipolar Couplings 319Malene Ringkjøbing Jensen, Valery Ozenne, Loic SalmonGabrielle Nodet, Phineus Markwick, Pau Bernado´ and MartinBlackledge9.1 Introduction 3199.2 Ensemble Descriptions of Unfolded Proteins 3209.3 Experimental Techniques for the Characterisation of IDPs3209.4 NMR Spectroscopy of Intrinsically Disordered Proteins3219.5 Residual Dipolar Couplings 3239.6 Conclusions 340References 340Index 347