Crystallographic Methods and Protocols

1. Front Matter

   Pages i-xii

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2. Introduction

   • Mark R. Sanderson

   Pages 1-21

3. Overexpression, Isolation, and Crystallization of Proteins

   • Jane V. Skelly, C. Bernadette Madden

   Pages 23-53

4. Preliminary Characterization of Crystals

   • Sherin S. Abdel-Meguid, David Jeruzalmi, Mark R. Sanderson

   Pages 55-86

5. Modern Methods for Rapid X-Ray Diffraction Data Collection from Crystals of Macromolecules

   • Elspeth F. Garman

   Pages 87-126

6. Use of Multiple-Wavelength Anomalous Diffraction Measurements in Ab Initio Phase Determination for Macromolecular Structures

   • H. M. Krishna Murthy

   Pages 127-151

7. Structure Determination Using Isomorphous Replacement

   • Sherin S. Abdel-Meguid

   Pages 153-171

8. Molecular Replacement Using Known Structural Information

   • Ian J. Tickle, Huub P. C. Driessen

   Pages 173-203

9. Density Modification in X-Ray Crystallography

   • Alberto D. Podjarny, Bernard Rees, Alexandre G. Urzhumtsev

   Pages 205-226

10. Refinement of Protein and Nucleic Acid Structures

    • Eric Westhof, Philippe Dumas

    Pages 227-244

11. Recent Developments for Crystallographic Refinement of Macromolecules

    • Axel T. Brünger

    Pages 245-266

12. The Crystallization and Structure Analysis of Oligonucleotide Sequences

    • Stephen Neidle

    Pages 267-292

13. Crystallography in the Study of Protein-DNA Interactions

    • David G. Brown, Paul S. Freemont

    Pages 293-318

14. Virus Crystallography

    • Elizabeth Fry, Derek Logan, David Stuart

    Pages 319-363

15. Crystallization and Structure Analysis of Membrane Proteins

    • Richard Newman

    Pages 365-387

16. Back Matter

    Pages 389-394

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The volumes in the series, Methods in Molecular Biology, are conceived with the biochemist and molecular biologist in mind. The present book, Crystallographic Methods andProtocols, concentrates on the use of X-ray crystallography to solve the detailed three­ dimensional structuresofproteins, nucleic acids, andtheir complexes. Such a structure determination is a major undertaking, demands expertise in a range of skills, and requires considerable resources. The biologically trained worker will probably first become involved when identifying an important scientific problem whose solution would benefit from a full structure. The protein or nucleic acid at issue must be sequenced and prepared to high purity in appropriate quantities, probably by either chemical synthesis for nucleic acids or genetic engineering for proteins. CrystallographicMethods andPro­ tocols aims to give biologically trained workers an insight into the techniques used to crystallize their proteins, obtain the raw X-ray data, and solve and refine the structure. The aim ofa crystal structure determination is to provide infor­ mation that will solve biologically relevant problems; that process normally requires a high resolution structure. The preparationofsuit­ able crystals for a high resolution structure remains the major bottle­ neck in structure determination, and the effective application of appropriate genetic engineering and biochemical techniques in the initial stages pays ahandsome dividend later. The productionofsuit­ able proteins by recombinant methods and the preparationofcrystal­ line derivatives are covered by Skelly and Madden in Chapter 2. Before beginning the data collection required for a high resolution structure, a preliminary characterizationofthe crystals is carried out.

...fills an important niche...delivers what it promises: methods and protocols; very basic and down-to-earth, but all the more useful...I warmly recommend this book to newcomers to the field as a hands-on guide to many of the varied activities a macromolecular crystallographer faces.-FEBS Letters

• National Institute for Biological Standards and Control, Potters Bar, UK

  Christopher Jones, Barbara Mulloy

• Department of Biophysics, King’s College, London, UK

  Mark R. Sanderson

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