pyrimidinones and pyrene

The structure of sarcoplasmic reticulum membranes was studied in the presence of modeled transmembrane Ca2+ gradient corresponding to the status of Ca2+ depot at different stages of the muscle contraction-relaxation cycle in health and disease. Various sites of the membrane were characterized using spectral analysis of tryptophan, pyrene, and merocyanine-540 fluorescence without evaluating specific changes in the molecules of membrane components (Ca2+ -ATPase, ryanodine receptor, and lipids). The transmembrane Ca2+ gradient modulates the protein-lipid interactions and structural characteristics of the membrane. The proposed model can be used for studies of the effects of pharmacologically active substances and endogenous regulators.

Topics: Animals; Calcium; Calcium-Transporting ATPases; Fluorescence; Intracellular Membranes; Membrane Lipids; Models, Theoretical; Protein Binding; Pyrenes; Pyrimidinones; Rabbits; Ryanodine Receptor Calcium Release Channel; Sarcoplasmic Reticulum; Spectrometry, Fluorescence; Tryptophan

The properties of liquid-ordered, solid-ordered, and liquid-disordered phases were investigated by steady-state fluorescence spectroscopy in liposomes composed of mixtures of dipalmitoylphosphatidylcholine and cholesterol (0-40 mol %) as a function of temperature (24-51 degrees C). The fluorescent probes used (bis-pyrene, nystatin, prodan, and merocyanine) were chosen because they differ in the location they occupy in the membrane and in the types of properties they sense. Comparison of phase diagrams with contour plots of the fluorescence data suggested that bis-pyrene is sensitive primarily to lipid order. In contrast, nystatin fluorescence intensity responded to changes in lipid fluidity. The shape of the prodan emission spectrum detected both liquid-solid and order-disorder transitions in the phase diagram. Merocyanine's behavior was more complex. First, it was more sensitive than any of the other probes to the membrane pretransition that occurs in the absence of cholesterol. Second, regardless of whether emission intensity, anisotropy, or spectral shape was observed, the probe appeared to distinguish two types of liquid-ordered phases, one with tightly packed lipids and one in which the apparent spacing among lipids was increased. The prodan data supported these results by displaying modest versions of these two observations. Together, the results identify eight regions within the phase diagram of distinguishable combinations of these physical properties. As an example of how this combined analysis can be applied to biological membranes, human erythrocytes were treated similarly. Temperature variation at constant cholesterol content revealed three of the eight combinations identified in our analysis of liposomes.

Topics: 1,2-Dipalmitoylphosphatidylcholine; 2-Naphthylamine; Anisotropy; Biophysics; Cholesterol; Fluorescent Dyes; Lipids; Microscopy, Fluorescence; Nystatin; Phospholipids; Pyrenes; Pyrimidinones; Spectrometry, Fluorescence; Temperature