It is proposed to determine the effects of ethanol on protein-lipid interactions in membranes and how lipids modified as a result of chronic ethanol ingestion might modulate the effects. The first aspect we propose to investigate is the binding and penetration of proteins which spontaneously incorporate into membranes. We will determine how this is affected by ethanol and whether is differs for lipids from membranes of ethanol tolerant animals as compared to controls using cytochrome b5 and apocytochrome C. The second aspect of protein-lipid interaction. This region has been difficult to study in the past but we have developed a new approach which in effect spectroscopically isolates this region, using time resolved fluorescence. We propose to use this approach to investigate the effect of ethanol perturbation on this region and whether it differs for different proteins. We will use cytochrome b5, apocytochrome C, gramicidin and glycophorin, as relatively simple representative membrane proteins. We will also determine whether the protein-lipid interfacial region differs in protein reconstituted with lipids from ethanol-tolerant animals. The functional significance of ethanol perturbation of the protein-lipid interfacial region will be explored using skeletal muscle sarcoplasmic reticulum calcium ATP'ase.
