The secondary structure of calcium pump protein in light sarcoplasmic reticulum and reconstituted in a single lipid component as determined by Raman spectroscopy

Raman spectra have been measured of the following samples: active calcium pump protein in light sarcoplasmic reticulum (SR) membranes, lipids extracted from light SR membranes, active calcium pump protein reconstituted in dielaidoylphosphatidylcholine (DEPC), and pure DEPC. The spectra of native SR lipids and of pure DEPC are different, and yet when these spectra are subtracted from the spectra of the respective protein-lipid complexes, the resulting amide I spectra of the calcium pump protein are the same, indicating that appropriate criteria have been chosen for subtraction of the spectrum of a lipid. This spectrum has been analyzed for secondary structure with the following results. The SR calcium pump protein contains 51 +/- 5% helix, in agreement with a prediction of secondary structure obtained from an analysis of the sequence, and 21 +/- 4% beta-strand. In addition, the presence of protein broadens and lowers the main melting transition of DEPC.