Lipid-protein interaction in the glycophorin-dipalmitoylphosphatidylcholine system: Raman spectroscopic investigation.

Raman spectra have been recorded as a function of temperature for lipid-protein complexes of glycophorin isolated from erythrocyte membranes reconstituted with dipalmitoylphosphatidylcholine (DPPC) and its chain perdeuterated analogue ([(2)H(62)]DPPC). The conformation of the phospholipid hydrocarbon chains in the vicinity of protein is drastically altered from that in pure lipid dispersions. Analysis of the chain C-(2)H stretching vibrations for complexes of [(2)H(62)]-DPPC-glycophorin shows that at lipid:protein mole ratios of 125:1, a broad melting event occurs that is not observable by calorimetric techniques. The midpoint occurs at temperatures about 15 degrees C below that of the gel/liquid crystal phase transition for [(2)H(62)]DPPC in multilamellar dispersions. The same number of gauche rotamers form in the phospholipid hydrocarbon chains during the melting process as in the phase transition of the unperturbed molecule. Analysis of the C-H stretching region of the Raman spectrum in DPPC-glycophorin complexes indicates that lateral interactions between phospholipid chains in the complex are reduced so that interchain vibrational coupling is minimized. The observed differences between the Raman melting curves and the calorimetric endothermic transitions arise because different populations of phospholipid molecules are sampled in the two experiments. The advantages of Raman spectroscopy for the study of lipid-protein interaction are demonstrated in the current work. Implications for the structure of the lipid in the immediate vicinity of membrane protein are discussed.