Protein NMR spectroscopy : practical techniques and applications / edited by Lu-Yun Lian, Gordon Roberts.

Format:

Book

Contributor:

Lian, Lu-Yun.

Roberts, G. C. K. (Gordon Carl Kenmure)

Language:

English

Subjects (All):

Proteins--Analysis.

Proteins.

Nuclear magnetic resonance spectroscopy.

Physical Description:

1 online resource (380 p.)

Edition:

1st ed.

Place of Publication:

Hoboken, N.J. : Wiley, 2011.

Language Note:

English

Summary:

"Nuclear Magnetic Resonance (NMR) spectroscopy is a powerful technique in structural biology for obtaining high resolution 3-D structures of proteins, second only, and complementary to X-ray crystallography. Molecules are studied in solution, where conditions are closer to what is found in the cell. It is the primary technique used to obtain information on intrinsically disordered (unfolded) proteins, since these proteins will not crystallize easily.The aim of this book is to provide the newcomer to NMR techniques with practical guidance on how to choose the right experiment, how to carry out the experiment, and how to analyse the resulting spectra. Those who are familiar with the chemical applications of NMR will also find it helpful in describing the special requirements of proteins"-- Provided by publisher.

Contents:

Protein NMR Spectroscopy: Practical Techniques and Applications; Contents; List of Contributors; Introduction; References; 1 Sample Preparation, Data Collection and Processing; 1.1 Introduction; 1.2 Sample Preparation; 1.2.1 Initial Considerations; 1.2.2 Additives; 1.2.3 Sample Conditions; 1.2.4 Special Cases; 1.2.5 NMR Sample Tubes; 1.2.5.1 3 mm Tubes; 1.3 Data Collection; 1.3.1 Locking; 1.3.2 Tuning; 1.3.3 Shimming; 1.3.4 Calibrating Pulses; 1.3.5 Acquisition Parameters; 1.3.6 Fast Acquisition Methods; 1.4 Data Processing; References; 2 Isotope Labelling; 2.1 Introduction

2.2 Production Methods for Isotopically Labelled Proteins2.2.1 Recombinant Protein Expression in Living Organisms; 2.2.1.1 Escherichia coli; 2.2.1.2 Yeast Cells; 2.2.1.3 Other Host Cells; 2.2.2 Cell-Free Synthesis; Protocol 1: Preparation of the Amino Acid Free S30 Extract; Protocol 2: Cell-Free Reaction on a Small Scale; 2.3 Uniform Isotope Labelling of Proteins; 2.3.1 Uniform 15N Labelling; 2.3.2 Uniform 13C, 15N Labelling; 2.3.3 2H Labelling; 2.4 Selective Isotope Labelling of Proteins; 2.4.1 Amino Acid Type-Selective Labelling; 2.4.2 Reverse Labelling; 2.4.3 Stereo-Selective Labelling

2.5 Segmental Labelling2.6 SAIL Methods; 2.6.1 Concept of SAIL; 2.6.2 Practical Procedure for the SAIL Method; Protocol 3: Production of SAIL Proteins by the E. coli Cell-Free Method; 2.6.3 Residue-Selective SAIL Method; Protocol 4: Optimisation of the Amount of SAIL Amino Acids for the Production of Calmodulin Selectively Labelled by SAIL Phenylalanine; 2.7 Concluding Remarks; Acknowledgements; References; 3 Resonance Assignments; 3.1 Introduction; 3.2 Resonance Assignment of Unlabelled Proteins; 3.2.1 Spin System Assignments; 3.2.2 Sequence-Specific Assignments; 3.2.3 Possible Difficulties

3.3 15N-Edited Experiments3.4 Triple Resonance; 3.4.1 3D Triple Resonance; 3.4.1.1 Identification of Spin Systems; 3.4.1.2 Sequential Assignment; 3.4.1.3 Proline Residues; 3.4.2 4D Triple Resonance; 3.4.3 Computer-Assisted Backbone Assignments; 3.4.4 Unstructured Proteins; 3.4.5 Large Proteins; 3.5 Side-Chain Assignments; References; 4 Measurement of Structural Restraints; 4.1 Introduction; 4.2 NOE-Based Distance Restraints; 4.2.1 Physical Background; 4.2.2 NMR Experiments for Measuring the NOE; 4.2.3 Set-up of NOESY Experiments; 4.2.3.1 Estimation of T2s; Recipe 4.1: 1-1 Echo Experiment

Recipe 4.2: Set-up of Optimal Acquisition TimesRecipe 4.3: Set-up of a 3D 15N-Edited NOESY Experiment (Figure 4.2a); Recipe 4.4: Set-up of a 3D 13C-Edited NOESY Experiment; 4.2.4 Deriving Structural Information from NOE Cross-peaks; Recipe 4.5: Extraction of Distances Using Classes; Recipe 4.6: Extraction of Distances Using the Two-Spin Approximation; 4.2.5 Information Content of NOE Restraints; 4.3 Dihedral Restraints Derived from J-Couplings; 4.3.1 Physical Background; 4.3.2 NMR Experiments for Measuring J-Couplings; Recipe 4.7: E.COSY Experiment; Recipe 4.8: Quantitative J-Correlation

4.3.3 Deriving Structural Information from J-Couplings

Notes:

Description based upon print version of record.

Includes bibliographical references and index.

ISBN:

9786613177780

9781119972822

1119972825

9781283177788

1283177781

9781119972006

1119972000

9781119972013

1119972019

OCLC:

739118493