Effect of phospholipid substitution on the mobility of protein-bound spin labels in sarcoplasmic reticulum.
The influence of phospholipid environment upon the mobility of spin labels covalently bound to the Ca2+-transport ATPase (ATP phosphohydrolase [EC 3.6.1.3]) was studied by electron spin resonance spectroscopy in native and reconstituted sarcoplasmic reticulum membranes. Fragmented sarcoplasmic reticulum of rabbit skeletal muscle was covalently labeled with maleimide spin-labels of different chain length or with 4-(2-iodoacetamido)-2,2,6,6-tetramethylpiperidinooxyl, and the phospholipids were exchanged for dipalmitoylphosphatidylcholine or dioleoylphosphatidylcholine. With short-chain maleimide or iodoacetamide spin labels, the spectrum of the protein-bound label reflected the change in microenvironment caused by replacement of endogenous phospholipids with dipalmitoylphosphatidylcholine as a decrease in mobility. In contrast, after labeling with long-chain maleimide derivatives, there were no noticeable differences in the spectra before and after substitution with dipalmitophatidylcholine. Replacement of endogenous phospholipids with dioleoylphosphatidylcholine did not affect the spectra. The data indicate that increased viscosity in the environment of Ca2+-transport ATPase produced by replacement of sarcoplasmic reticulum lipids with dipalmitoylphosphatidylcholine reduces the mobility of short-chain maleimide spin labels covalently attached to the Ca2+-transport ATPase polypeptide.
D000818:Animals; D000252:Calcium-Transporting ATPases / Q000378:metabolism*; D004578:Electron Spin Resonance Spectroscopy; D007461:Iodoacetates / Q000494:pharmacology; D008301:Maleimides / Q000494:pharmacology; D010743:Phospholipids / Q000378:metabolism*; D011663:Pulmonary Surfactants / Q000494:pharmacology; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000378:metabolism*; D013113:Spin Labels; D013696:Temperature
D007461:Iodoacetates; D008301:Maleimides; D010743:Phospholipids; D011663:Pulmonary Surfactants; D013113:Spin Labels; D000252:Calcium-Transporting ATPases