Flavoprotein Protocols

1. Front Matter

   Pages i-xii

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2. UV-Visible Spectroscopy as a Tool to Study Flavoproteins

   • Peter Macheroux

   Pages 1-7

3. Identifying and Quantitating FAD and FMN in Simple and in Iron-Sulfur-Containing Flavoproteins

   • Alessandro Aliverti, Bruno Curti, Maria A. Vanoni

   Pages 9-23

4. Fluorescence Analysis of Flavoproteins

   • Andrew W. Munro, Michael A. Noble

   Pages 25-48

5. Potentiometric Measurement of Oxidation-Reduction Potentials

   • Stephen G. Mayhew

   Pages 49-59

6. Flavoprotein Kinetics

   • Willem J. H. van Berkel, Jacques A. E. Benen, Michel H. M. Eppink, Marco W. Fraaije

   Pages 61-85

7. Application of Freeze Quenching to the Study of Rapid Reactions in Flavoproteins

   • W. J. Ingledew

   Pages 87-95

8. Application of Electron Spin Resonance (ESR) for Detection and Characterization of Flavoprotein Semiquinones

   • Marat B. Murataliev

   Pages 97-110

9. Circular Dichroism Studies of Flavoproteins

   • Andrew W. Munro, Sharon M. Kelly, Nicholas C. Price

   Pages 111-123

10. Vibrational Spectroscopy of Flavoproteins

    • Iain D. G. Macdonald

    Pages 125-138

11. NMR of Flavoproteins

    • Jacques Vervoort, Marco Hefti

    Pages 139-147

12. Flavoprotein Resolution and Reconstitution

    • Florence Lederer, Heinz Rüterjans, Garrit Fleischmann

    Pages 149-155

13. Flavoenzyme Structure and Function

    • Dale Edmondson, Sandro Ghisla

    Pages 157-179

14. Identification of Covalent Flavoproteins and Analysis of the Covalent Link

    • Nigel S. Scrutton

    Pages 181-193

15. Flavin Synthesis and Incorporation into Synthetic Peptides

    • R. E. Sharp, P. L. Dutton

    Pages 195-206

16. Computational Methods in Flavin Research

    • Lars Ridder, Hans Zuilhof, Jacques Vervoort, Ivonne M. C. M. Rietjens

    Pages 207-228

17. Flavins and Flavoenzymes in Diagnosis and Therapy

    • Katja Becker, Markus Schirmer, Stefan Kanzok, R. H. Schirmer

    Pages 229-245

18. Back Matter

    Pages 247-256

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As a scientist with an interest in proteins you will, at some time in your career, isolate an enzyme that turns out to be yellow—or perhaps you already have. Alternatively, you may identify a polypeptide sequence that is related to known flavin-containing proteins. This may, or may not, be your first encounter with flavoproteins. However, even if you are an old hand in the field, you may not have exploited the full range of experimental approaches applicable to the study of flavoproteins. We hope that Flavoprotein Protocols will encourage you to do so. In this volume we have sought to bring together a range of experimental methods of value to researchers with an interest in flavoproteins, whether or not these researchers have experience in this area. A broad range of techniques, from the everyday to the more specialized, is described by scientists who are experts in their fields and who have ext- sive practical experience with flavoproteins. The wide range of approaches, from wet chemistry to dry computation, has, as a consequence, demanded a range of formats. Where appropriate (particularly for analytical methods) the protocol described is laid out in easy-to-follow steps. In other cases (e. g. , the more advanced spectroscopies and computational methods) it is far more apt to describe the general approach and relevance of the methods. We hope this wide-ranging approach will sow the seeds of many future collaborations - tween laboratories and further our knowledge and understanding of how f- voproteins work.

"A detailed analysis of flavoproteins will be of value for investigators working with flavin enzymes whether studying mechanisms or searching for inhibitors. Chapters on chemical syntheses and the use of analogs provide useful information on approaches to both of these areas.."-Doodys Health Sciences Book Review Journal

• University of Edinburgh, Edinburgh, UK

  Stephen K. Chapman, Graeme A. Reid

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