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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.
Sommario
Intrinsic Fluorescence of Proteins.- Spectral Enhancement of Proteins by in vivo Incorporation of Tryptophan Analogues.- Room Temperature Tryptophan Phosphorescence as a Probe of Structural and Dynamic Properties of Proteins.- Azurins and Their Site-Directed Mutants.- Barnase: Fluorescence Analysis of A Three Tryptophan Protein.- Fluorescence Study of the DsbA Protein from Escherichia Coli.- The Conformational Flexibility of Domain III of Annexin V is Modulated by Calcium, pH and Binding to Membrane/Water Interfaces.- Tryptophan Calmodulin Mutants.- Luminescence Studies with trp Aporepressor and Its Single Tryptophan Mutants.- Heme-Protein Fluorescence.- Conformation of Troponin Subunits and Their Complexes from Striated Muscle.- Fluorescence of Extreme Thermophilic Proteins.
Info autore
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.
