Protein Fluorescence

1. Front Matter

   Pages i-xxi

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2. Intrinsic Fluorescence of Proteins

   • Maurice R. Eftink

   Pages 1-15

3. Spectral Enhancement of Proteins by in vivo Incorporation of Tryptophan Analogues

   • J. B. Alexander Ross, Elena Rusinova, Linda A. Luck, Kenneth W. Rousslang

   Pages 17-42

4. Room Temperature Tryptophan Phosphorescence as a Probe of Structural and Dynamic Properties of Proteins

   • Vinod Subramaniam, Duncan G. Steel, Ari Gafni

   Pages 43-65

5. Azurins and Their Site-Directed Mutants

   • Giampiero Mei, Nicola Rosato, Alessandro Finazzi Agro

   Pages 67-81

6. Barnase: Fluorescence Analysis of A Three Tryptophan Protein

   • Yves Engelborghs, Alan Fersht

   Pages 83-101

7. Fluorescence Study of the DsbA Protein from Escherichia Coli

   • Alain Sillen, Jens Hennecke, Rudi Glockshuber, Yves Engelborghs

   Pages 103-121

8. The Conformational Flexibility of Domain III of Annexin V is Modulated by Calcium, pH and Binding to Membrane/Water Interfaces

   • Jacques Gallay, Jana Sopková, Michel Vincent

   Pages 123-173

9. Tryptophan Calmodulin Mutants

   • Jacques Haiech, Marie-Claude Kilhoffer

   Pages 175-209

10. Luminescence Studies with trp Aporepressor and Its Single Tryptophan Mutants

    • Maurice R. Eftink

    Pages 211-219

11. Heme-Protein Fluorescence

    • Rhoda Elison Hirsch

    Pages 221-255

12. Conformation of Troponin Subunits and Their Complexes from Striated Muscle

    • Herbert C. Cheung, Wen-Ji Dong

    Pages 257-283

13. Fluorescence of Extreme Thermophilic Proteins

    • Sabato D’Auria, Mosè Rossi, Ignacy Gryczynski, Joseph R. Lakowicz

    Pages 285-306

14. Back Matter

    Pages 307-310

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The intrinsic or natural fluorescence of proteins is perhaps the most complex area of biochemical fluorescence. Fortunately the fluorescent amino acids, phenylalanine, tyrosine and tryptophan are relatively rare in proteins. Tr- tophan is the dominant intrinsic fluorophore and is present at about one mole % in protein. As a result most proteins contain several tryptophan residues and even more tyrosine residues. The emission of each residue is affected by several excited state processes including spectral relaxation, proton loss for tyrosine, rotational motions and the presence of nearby quenching groups on the protein. Additionally, the tyrosine and tryptophan residues can interact with each other by resonance energy transfer (RET) decreasing the tyrosine emission. In this sense a protein is similar to a three-particle or mul- particle problem in quantum mechanics where the interaction between particles precludes an exact description of the system. In comparison, it has been easier to interpret the fluorescence data from labeled proteins because the fluorophore density and locations could be controlled so the probes did not interact with each other. From the origins of biochemical fluorescence in the 1950s with Prof- sor G. Weber until the mid-1980s, intrinsic protein fluorescence was more qualitative than quantitative. An early report in 1976 by A. Grindvald and I. Z. Steinberg described protein intensity decays to be multi-exponential. Attempts to resolve these decays into the contributions of individual tryp- phan residues were mostly unsuccessful due to the difficulties in resolving closely spaced lifetimes.

• Calcium
• Phosphor
• escherichia coli
• fluorescence
• luminescence
• membrane
• mutant
• protein
• spectra
• spectroscopy
• temperature
• units
• water

• Center for Fluorescence Spectroscopy and Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore

  Joseph R. Lakowicz

Dr. J.R. Lakowicz is Professor of Biochemistry at the University of Maryland School of Medicine and Director of the Center for Fluorescence Spectroscopy. Dr. Lakowicz has published over 400 scientific articles, has edited numerous books, holds 16 issued patents, and is the sole author of the widely used text, Principles of Fluorescence Spectroscopy, also published by Kluwer Academic/Plenum Publishers, now in its Second Edition.

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