Temperature-induced transitions of porcine intestinal brush border membranes.
Thermotropic transitions of the membrane components in porcine intestinal brush border membranes were studied by means of fluorimetry using a fluorogenic thiol reagent, N-[7-dimethylamino-4-methylcoumarinyl]maleimide (DACM), and a lipophilic fluorescent probe, 1,6-diphenyl-1,3,5-hexatriene (DPH). 1. The reactivity of the sulfhydryl groups of the membrane proteins with DACM was dependent on temperature, with a transition point at about 33 degrees C. A conspicuous transition was also observed in the relation between temperature and the fluorescence intensity of DACM-labeled membranes at 35 degrees C. 2. Temperature dependence profiles of the solubilization of DPH in the membranes and of the fluorescence polarization of DPH-membrane complex suggested that the phase transition of the lipid from gel to liquid-crystalline state occurs over a temperature range of 30 to 35 degrees C. 3. Efficient fluorescence energy transfer was observed from tryptophan residues of the membrane proteins to DPH located in the lipid phase of the membranes, and its efficiency was extremely enhanced, dependent on temperature, above 35 degrees C. The intensity of the tryptophan fluorescence of the membrane proteins decreased with increasing temperature and a discontinuity was observed at about 33 degrees C. Based on these results, it may be concluded that there are co-operative interactions between proteins and lipids in the membranes and that the temperature-induced conformational changes of the membrane proteins are closely related to the dynamics of the hydrocarbon cores of the lipid.
D000818:Animals; D002462:Cell Membrane / Q000502:physiology*; D004735:Energy Transfer; D005454:Fluorescence Polarization; D007413:Intestinal Mucosa; D008560:Membrane Fluidity; D008563:Membrane Lipids; D008565:Membrane Proteins; D008871:Microvilli / Q000502:physiology*; D013050:Spectrometry, Fluorescence; D013552:Swine; D013696:Temperature; D014364:Tryptophan
D008563:Membrane Lipids; D008565:Membrane Proteins; D014364:Tryptophan