X-Ray Crystallography of Biomacromolecules A Practical Guide

by Messerschmidt, Albrecht

• ISBN13: 9783527313969
• ISBN10: 3527313966
• Edition: 1st
• Format: Hardcover
• Copyright: 2007-01-23
• Publisher: Wiley-Blackwell

Summary

Written by one of the most significant contributors to the progress of protein crystallography, this practical guide contains case studies, a troubleshooting section and pointers on data interpretation. It covers the theory, practice and latest achievements in x-ray crystallography, such that any researcher in structural biology will benefit from this extremely clearly written book.Part A covers the theoretical basis and such experimental techniques as principles of x-ray diffraction, solutions for the phase problem and time-resolved x-ray crystallography. Part B includes case studies for different kinds of x-ray crystal structure determination, such as the MIRAS and MAD techniques, molecular replacement, and the difference Fourier technique.

Author Biography

Professor Dr. Albrecht Messerschmidt is group leader at the Max Planck Institute for Biochemistry, Marinsried, Germany.
His main reserach interest is in the determination of X-ray structures of metalloproteins and -enzymes, PLP-dependent enzymes, CoA-transferases and other proteins. His group is part of the department of X-ray structure determination of proteins headed by nobel laureate Robert Huber.

Table of Contents

Ptreface

Principles and Methods

Introduction

Crystals and Symmetry

Protein Solubility

Experimental Techniques

Crystallization Screenings

High-Throughput Crystallization, Imaging, and Analysis. References

Experimental Techniques

X-Ray Sources

Detectors

Crystal Mounting and Cooling

Data Collection Techniques. References

Principles of X-Ray Diffraction by a Crystal

Rational Mathematical Representation of Waves

Principles of X-Ray Diffraction by a Crystal. References

Diffraction Data Evaluation

Introductory Remarks

Geometric Principles in the Rotation Technique with Normal Flat Detector

Autoindexing of Oscillation Images

Beam Divergence, Mosaicity, and Partiality

Integration of Diffraction Spots

Post-Refinement, Scaling, and Averaging of Diffraction Data. References

Methods for Solving the Phase Problem

Isomorphous Replacement

Anomalous Scattering

Determination of Heavy-Atom Positions

Phase Calculation

Patterson Search Methods (Molecular Replacement). References

Phase Improvement by Density Modification and Phase Combination

Introduction

Solvent Flattening

Histogram Matching

Molecular Averaging

Sayre's Equation

Atomization

Phase Combination

Difference Fourier Technique. References

Model Building and Refinement

Model Building

Crystallographic Refinement

Verification and Accuracy of Structure Determination. References

Crystal Structure Determination of the Time-Course of Reactions and of Unstable Species

Introduction

Triggering Methods

Trapping Methods

Laue Diffraction. References

Structural Genomics

Introduction

Target Selection

Production of Recombinant Proteins

Aspects of Automation. References

Practical Examples. Introductory Remarks

Data Evaluation

Autoindexing, Refinement of Cell Parameters, and Reflection Integration

Scaling of Intensity Diffraction Data

A Complex Example of Space Group Determination. References

Determination of Anomalous Scatterer or Heavy Atom Positions

Application of Direct Methods

Vector Verification Methods

Comparison of the Results from SnB and RSPS. References

MIRAS and MAD Phasing with the Program SHARP

MAD Phasing with the Program SHARP for 4-BUDH. References

Molecular Replacement

Phase Determination of PKC-iota with Program Molrep. References

Averaging about Non-Crystallographic Symmetry (NCS) for 4-BUDH

Determination of NCS Operators for 4-BUDH

Electron Density Map Averaging for 4-BUDH. References

Model Building and More

A Very Personal Short Introduction to the Computer Graphics Modeling Program "O"

Introduction of the Four Fe-Sites per Fe-S-Cluster and New SHARP-Phasing for 4-BUDH

Crystallographic Refinement and Final Steps

References

Subject Index

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