Roberts / Lian Protein NMR Spectroscopy

Practical Techniques and Applications
1. Auflage 2011
ISBN: 978-1-119-97282-2
Verlag: John Wiley & Sons
Format: EPUB
Kopierschutz: Adobe DRM (»Systemvoraussetzungen)

E-Book, Englisch, 384 Seiten, E-Book

Nuclear Magnetic Resonance (NMR) spectroscopy, a physicalphenomenon based upon the magnetic properties of certain atomicnuclei, has found a wide range of applications in life sciencesover recent decades. The dramatic advances in NMR techniques haveled to corresponding advances in the ability of NMR to studystructure, dynamics and interactions of biological macromoleculesin solution under close to physiological conditions. This volumefocuses on the use of NMR to study proteins.
NMR can be used to determine detailed three-dimensionalstructures of proteins in solution. Furthermore, it providesinformation about conformational or chemical exchange, internalmobility and dynamics at timescales varying from pcoseconds toseconds. It is the primary technique used to obtain information onintrinsically disordered (unfolded) proteins, since these proteinswill not crystallize easily. NMR is also a very powerful method forthe study of interactions of protein with other molecules, whethersmall molecules (including drugs), nuclear acids or otherproteins.
This up-to-date volume covers NMR techiniques and theirapplication to proteins, with a focus on practical details. Thisbook will provide a newcomer to NMR with the practical guidance inorder to carry out successful experiments with proteins and toanalyze the resulting spectra. Those who are familiar with thechemical applications of NMR will also find is useful inunderstanding the special requirements of protien NMR.

Autoren/Hrsg.

Roberts, Gordon

Lian, Lu-Yun

Inhaltsverzeichnis

List of Contributors xiii
Introduction 1
Lu-Yun Lian and Gordon Roberts
References 4
1 Sample Preparation, Data Collection and Processing5
Frederick W. Muskett
1.1 Introduction 5
1.2 Sample Preparation 5
1.3 Data Collection 11
1.4 Data Processing 17
2 Isotope Labelling 23
Mitsuhiro Takeda and Masatsune Kainosho
2.1 Introduction 23
2.2 Production Methods for Isotopically Labelled Proteins 24
2.3 Uniform Isotope Labelling of Proteins 29
2.4 Selective Isotope Labelling of Proteins 32
2.5 Segmental Labelling 37
2.6 SAIL Methods 38
2.7 Concluding Remarks 45
3 Resonance Assignments 55
Lu-Yun Lian and Igor L. Barsukov
3.1 Introduction 55
3.2 Resonance Assignment of Unlabelled Proteins 56
3.3 15N-Edited Experiments 60
3.4 Triple Resonance 62
3.5 Side-Chain Assignments 77
4 Measurement of Structural Restraints 83
Geerten W. Vuister, Nico Tjandra, Yang Shen, Alex Grishaev andStephan Grzesiek
4.1 Introduction 83
4.2 NOE-Based Distance Restraints 84
4.3 Dihedral Restraints Derived from J-Couplings 96
4.4 Hydrogen Bond Restraints 103
4.5 Orientational Restraints 107
4.6 Chemical Shift Structural Restraints 129
4.7 Solution Scattering Restraints 137
5 Calculation of Structures from NMR Restraints 159
Peter Guntert
5.1 Introduction 159
5.2 Historical Development 161
5.3 Structure Calculation Algorithms 164
5.4 Automated NOE Assignment 173
5.5 Nonclassical Approaches 178
5.6 Fully Automated Structure Analysis 181
6 Paramagnetic Tools in Protein NMR 193
Peter H.J. Keizers and Marcellus Ubbink
6.1 Introduction 193
6.2 Types of Restraints 194
6.3 What Metals to Use? 200
6.4 Paramagnetic Probes 203
6.5 Examples 209
6.6 Conclusions and Perspective 212
7 Structural and Dynamic Information on Ligand Binding221
Gordon Roberts
7.1 Introduction 221
7.2 Fundamentals of Exchange Effects on NMR Spectra 222
7.3 Measurement of Equilibrium and Rate Constants 229
7.4 Detecting Binding - NMR Screening 238
7.5 Mechanistic Information 241
7.6 Structural Information 246
8 Macromolecular Complexes 269
Paul C. Driscoll
8.1 Introduction 269
8.2 Spectral Simplification through Differential IsotopeLabelling 270
8.3 Basic NMR Characterisation of Complexes 273
8.4 3D Structure Determination of MacromolecularProtein-Ligand Complexes 277
8.5 Literature Examples 297
9 Studying Partially Folded and Intrinsically DisorderedProteins Using NMR Residual Dipolar Couplings 319
Malene Ringkjøbing Jensen, Valery Ozenne, Loic Salmon,Gabrielle Nodet, Phineus Markwick, Pau Bernado´ and MartinBlackledge
9.1 Introduction 319
9.2 Ensemble Descriptions of Unfolded Proteins 320
9.3 Experimental Techniques for the Characterisation of IDPs320
9.4 NMR Spectroscopy of Intrinsically Disordered Proteins321
9.5 Residual Dipolar Couplings 323
9.6 Conclusions 340
References 340
Index 347