Librational motions of membrane-embedded Ca2+-ATPase of sarcoplasmic reticulum labeled with fluorescein isothiocyanate.
Continuing our investigation of the relationships between internal motions and functional properties of soluble and membrane-bound proteins we have explored the lifetimes and correlation times associated with the fluorescence emission of fluorescein-labeled Ca2+-dependent ATPase of sarcoplasmic reticulum. The emission was characterized by two lifetime components near 1.8 and 4.1 ns, probably due to exposure of the probe to environments of different polarities. The time-dependent anisotropy showed the presence of two correlation times near 0.8 and 6.6 ns. The shorter correlation time was due to motions of the probe around its point of attachment on the surface of the protein. The longer correlation time indicated the presence of internal motions of the protein. Both lifetimes and correlation times were insensitive to temperature between 2 and 10 degrees C. They were also insensitive to addition and removal of 100 microM free Ca2+.
D000818:Animals; D000252:Calcium-Transporting ATPases / Q000378:metabolism*; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007700:Kinetics; D009132:Muscles / Q000201:enzymology; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000201:enzymology*; D013861:Thiocyanates
D005452:Fluoresceins; D005456:Fluorescent Dyes; D013861:Thiocyanates; D000252:Calcium-Transporting ATPases; D016650:Fluorescein-5-isothiocyanate