Structural and functional changes occurring in sarcoplasmic reticulum vesicles following exposure to low concentrations (5--7%, v/v) of diethyl ether in aqueous media, were studied by electron microscopy and by kinetic measurements of Ca2+ transport and ATPase activity. Electron microscopy of thin sectioned and freeze-fractured sarcoplasmic reticulum vesicles provided detailed resolution of Ca-ATPase amphiphilic molecules displaying 'lollipop' portions on the outer surface of the vesicle, and non-polar moieties penetrating the membrane's hydrophobic interior. This asymmetric disposition of ATPase molecules was disrupted in vesicles exposed to ether and then centrifuged and/or resuspended in aqueous media. Such vesicles had a tendency to undergo fragmentation, and the distribution of ATPase molecules was markedly altered. The continuous fuzzy layer of lollipops became discontinuous, and the intramembranous particles became randomly distributed over both the concave and the convex freeze-fracture membrane faces. Functionally, the vesicles lost their ability to accumulate calcium in the presence of ATP, although high rates of ATPase activity were maintained. Vesicles which were simply exposed to ether, without being subjected to centrifugation and/or homogenization, did not appear altered ultrastructurally, and retained their ability to accumulate calcium. In fact, the enzyme turnover and the maximal levels of calcium uptake were increased. It is concluded that diethyl ether interferes with lipid-lipid and protein-lipid interactions in the sarcoplasmic reticulum vesicle membrane, thereby facilitating molecular motions which may be a limiting factor in the transport mechanism. On the other hand, these weakened interactions permit structural denaturation and loss of the ability to maintain a transmembrane Ca2+ gradient when the vesicles are subjected to mechanical perturbations which are harmless in the absence of ether.