Electron microscopic analysis of two-dimensional crystals of the Ca2+-transport ATPase--a freeze-fracture study.

Two distinct forms of Ca2+-ATPase crystals have been analysed in sarcoplasmic reticulum (SR) membranes. The E1-type crystals, induced by Ca2+ or lanthanide ions, consist of single chains of ATPase monomers, and the E2-type crystals, induced by vanadate ions, consist of dimer chains. Using improved freeze-fracture techniques we have obtained high-resolution images of complementary surface replicas of SR membranes containing these crystal forms. In E1 crystals, the concave fracture (P) faces display obliquely oriented rows of intramembrane particles (IMPs) spaced at congruent to 6-7 nm along both crystal axes, while the convex fracture (E) faces show corresponding rows of pits. In E2 crystals, regular arrays of oblique parallel ridges with spacing of congruent to 10.5-11 nm appear on the P-faces and complementary grooves or furrows on the E-faces. In many instances the ridges break up into elongated particles repeating every 5.5 nm. When the direction of the shadow is almost parallel to the axis of the ridges, these 9.5 nm particles can be resolved into two domains, which represent intramembranous contacts between the two monomers of the two adjacent dimer chains. Complementary grooves on the E-faces can also be resolved into rows of pits complementary to the particles of the ridges on the P-faces. In the control SR membranes, randomly dispersed IMPs and corresponding pits are observed on the P- and E-faces, respectively. The data suggest that transport of Ca2+ involves significant structural changes of the enzyme molecule, reflected in the ATPase-ATPase interactions both on the cytoplasmic surface and in the lipid bilayer.