Red-edge excitation of fluorescence and dynamic properties of proteins and membranes. 1984

J R Lakowicz, and S Keating-Nakamoto

In moderately polar and viscous solvents, the emission spectra of fluorophores often shift to longer wavelengths as the excitation wavelength is increased toward the long-wavelength (red) side of the absorption. Red shifts occur because long-wavelength excitation results in photoselection of those fluorophores which are interacting most strongly with the polar solvent molecules. The observation of excitation red shifts requires that these enhanced dipole-dipole interactions are maintained in the photoselected population during the lifetime of the excited state. Consequently, the magnitude of the excitation red shifts depends upon the dynamic properties of the environment surrounding the fluorophore, as well as upon the solvent polarity and the sensitivity of the fluorophore to the polarity of the solvent. We used this phenomenon to investigate the dynamic properties of reference solvents, model membranes, and the protein apomyoglobin labeled with 6-(p-toluidinyl)-2-naphthalenesulfonic acid (TNS). The spectral shifts and lifetime data indicate that red-edge excitation results in the selective excitation of "solvent-relaxed" fluorophores. By comparison of the data obtained for TNS in solvents and bound to the macromolecules, one may estimate the relaxation rate of the environment. This comparison indicates rapid spectral relaxation of TNS bound to lipid vesicles and a somewhat slower relaxation around TNS bound to the heme site of apomyoglobin.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D008081 Liposomes Artificial, single or multilaminar vesicles (made from lecithins or other lipids) that are used for the delivery of a variety of biological molecules or molecular complexes to cells, for example, drug delivery and gene transfer. They are also used to study membranes and membrane proteins. Niosomes,Transferosomes,Ultradeformable Liposomes,Liposomes, Ultra-deformable,Liposome,Liposome, Ultra-deformable,Liposome, Ultradeformable,Liposomes, Ultra deformable,Liposomes, Ultradeformable,Niosome,Transferosome,Ultra-deformable Liposome,Ultra-deformable Liposomes,Ultradeformable Liposome
D008433 Mathematics The deductive study of shape, quantity, and dependence. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed) Mathematic
D009211 Myoglobin A conjugated protein which is the oxygen-transporting pigment of muscle. It is made up of one globin polypeptide chain and one heme group.
D009282 Naphthalenesulfonates A class of organic compounds that contains a naphthalene moiety linked to a sulfonic acid salt or ester.
D011485 Protein Binding The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments. Plasma Protein Binding Capacity,Binding, Protein
D011487 Protein Conformation The characteristic 3-dimensional shape of a protein, including the secondary, supersecondary (motifs), tertiary (domains) and quaternary structure of the peptide chain. PROTEIN STRUCTURE, QUATERNARY describes the conformation assumed by multimeric proteins (aggregates of more than one polypeptide chain). Conformation, Protein,Conformations, Protein,Protein Conformations
D005456 Fluorescent Dyes Chemicals that emit light after excitation by light. The wave length of the emitted light is usually longer than that of the incident light. Fluorochromes are substances that cause fluorescence in other substances, i.e., dyes used to mark or label other compounds with fluorescent tags. Flourescent Agent,Fluorescent Dye,Fluorescent Probe,Fluorescent Probes,Fluorochrome,Fluorochromes,Fluorogenic Substrates,Fluorescence Agents,Fluorescent Agents,Fluorogenic Substrate,Agents, Fluorescence,Agents, Fluorescent,Dyes, Fluorescent,Probes, Fluorescent,Substrates, Fluorogenic
D006418 Heme The color-furnishing portion of hemoglobin. It is found free in tissues and as the prosthetic group in many hemeproteins. Ferroprotoporphyrin,Protoheme,Haem,Heme b,Protoheme IX
D001059 Apoproteins The protein components of a number of complexes, such as enzymes (APOENZYMES), ferritin (APOFERRITINS), or lipoproteins (APOLIPOPROTEINS). Apoprotein

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