Two-dimensional diffusion of amphiphiles in phospholipid monolayers at the air-water interface.

Steady-state and time-resolved fluorescence spectroscopy has been used to examine lateral diffusion in dipalmitoyl-L-alpha-phosphatidylcholine (DPPC) and dimyristoyl-L-alpha-phosphatidylcholine (DMPC) monolayers at the air-water interface, by studying the fluorescence quenching of a pyrene-labeled phospholipid (pyrene-DPPE) by two amphiphilic quenchers. Steady-state fluorescence measurements revealed pyrene-DPPE to be homogeneously distributed in the DMPC lipid matrix for all measured surface pressures and only in the liquid-expanded (LE) phase of the DPPC monolayer. Time-resolved fluorescence decays for pyrene-DPPE in DMPC and DPPC (LE phase) in the absence of quencher were best described by a single-exponential function, also suggesting a homogeneous distribution of pyrene-DPPE within the monolayer films. Addition of quencher to the monolayer film produced nonexponential decay behavior, which is adequately described by the continuum theory of diffusion-controlled quenching in a two-dimensional environment. Steady-state fluorescence measurements yielded lateral diffusion coefficients significantly larger than those obtained from time-resolved data. The difference in these values was ascribed to the influence of static quenching in the case of the steady-state measurements. The lateral diffusion coefficients obtained in the DMPC monolayers were found to decrease with increasing surface pressure, reflecting a decrease in monolayer fluidity with compression.

Nonexponential fluorescence decay of aqueous tryptophan and two related peptides by picosecond spectroscopy.

Time-resolved fluorescence spectroscopy of tryptophan and two related dipeptides, tryptophylalanine and alanyltryptophan, has been carried out on the subnanosecond time scale by using picosecond exciting pulses at a wavelength of 264 nm. Detection was with an ultrafast streak camera coupled to an optical multichannel analyzer. The zwitterions of these molecules show a definite nonexponential fluorescence decay which can be analyzed in terms of two exponentials. The two decay rates increase strongly with increasing temperature, as does the weight of the faster component. In tryptophan at pH 11, where the amino group is deprotonated, there remains only a single temperature-dependent exponential. The results are interpreted in terms of two kinds of trapped conformers in the excited state that interconvert no quicker than the time scale of the fluorescence. A model is suggested in which the nonradiative processes in one conformer approximate those in the bare indole moiety. The nonradiative decay rate of the other conformer is substantially faster. It is believed that the process responsible for this fast decay is intramolecular electron transfer from the indole to the amino acid side chain. The predilection for this electron transfer depends on steric relationships as well as on the electron-attracting power of the carbonyl group. This picture is consistent with earlier fluorescence quantum yield results. In fact, a self-consistent picture emerges from the temporal and yield data that quantitatively explains most important facets of tryptophan photochemistry in aqueous solution.