Effects of organic solutes on the Raman spectra of barnacle muscle fibers.
The Raman spectra observed from barnacle muscle fibers are quite complex because the cytoplasm of these cells contains several proteins and solutes. An extraction procedure was used to separate organic solutes from the contractile proteins. Glycine, trimethylamine oxide, taurine, and alanine were found to contribute to the Raman spectra of barnacle muscle fibers, while spectra of lobster fibers reveal the presence of betaine in addition. We have observed that the increase in osmolarity of the intracellular fluid caused by the augmentation of the salinity of sea water (density, 1.023-1.030) in which the barnacles were kept, induces a reduction of intensity of the amide I band. To distinguish among the different parameters which are modified by the sea water salinity, observations were made on glycerinated barnacle muscle fibers. The reduction of intensity of the amide I band in the Raman spectra of glycerinated muscle fibers was also observed with the addition of taurine (0.08 M) in the external relaxing solution. Therefore, under these experimental conditions, the Raman scattering intensity in the amide I region assigned to the alpha-helix conformation (1645-1650 cm-1) is increased when the concentration of organic electrolytes is reduced. However, as no significant decrease of the scattering intensity in the 1660-1670 cm-1 region where the amide I bands of either beta-sheet or disordered conformations normally appear was observed, the increase of intensity of the amide I band centered at 1645 cm-1 is assigned to a change of orientation of alpha-helical segments of the myosin molecules. Our results suggest that organic solutes influence the position of the S-2 segments relative to the thick filaments.
Caillé
J P
JP
Département de biophysique, Faculté de médecine, Université de Sherbrooke, Québec, Canada.
Pigeon-Gosselin
M
M
Pézolet
M
M
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D003285:Contractile Proteins / Q000032:analysis; D066298:In Vitro Techniques; D009132:Muscles / Q000032:analysis*; D012996:Solutions; D013059:Spectrum Analysis, Raman / Q000379:methods; D001468:Thoracica
D000596:Amino Acids; D003285:Contractile Proteins; D012996:Solutions