No abstract

The structure of the E. coli F1 ATPase (ECF1) has been studied by a novel combination of two specimen preparation and image analysis techniques. The molecular outline of the ECF1 was determined by three-dimensional reconstruction of images of negatively stained two-dimensional crystals of ECF1. Internal features were revealed by analysis of single particles of ECF1, preserved in their native state in a thin layer of amorphous ice, and examined by cryoelectron microscopy. Various projections of the unstained ECF1 were interpreted consistently with the three-dimensional structure in negative stain, yielding a more informative description of the enzyme than otherwise possible. Results show that the ECF1 is a roughly spherical complex approximately 90-100 A in diameter. Six elongated protein densities (the alpha and beta subunits, each approximately 90 A X approximately 30 A in size) comprise its hexagonally modulated periphery. At the center of the ECF1 is an aqueous cavity which extends nearly or entirely through the length of the complex. A compact protein density, located at one end of the hexagonal barrel and closely associated with one of the peripheral subunits, partially obstructs the central cavity.

E. coli F1 F0 ATP synthase has been reconstituted into membranes and visualized by electron microscopy of unstained samples preserved in thin layers of amorphous ice. Unlike previous observations in negative stain, these specimens are not exposed to potentially denaturing or perturbing conditions, having been rapidly frozen from well‐defined conditions in which the enzyme is fully active. The structures visualized in views normal to the lipid bilayer clearly show the presence of a narrow stalk approx. 45 Å long, connecting the F1 to the membrane‐embedded F0.

A number of species of coccolithophorid phytoplankton precipitate calcite inside intracellular vesicles (coccolith vesicles). They can form vast blooms under certain conditions, and account for major fluxes of inorganic carbon (Ci) to the ocean floor. The functions of calcification have been debated for many years, and a role in carbon acquisition has been proposed by several workers. The precipitation of calcite from HCO3- involves the production of protons that can potentially be used to facilitate the use of external HCO3- as a photosynthetic substrate. For this function to be feasible, certain criteria must be met. HCO3- (rather than CO32–) should be the external substrate for calcification, photosynthesis should be facilitated by HCO3- in calcifying cells when CO2 availability is limiting, and the transport of Ci and Ca2+ to the site of calcification should be energetically and kinetically feasible. Considerable evidence exists for HCO3- as the substrate for calcification in coccolithophores. However, evidence for a direct role for calcification in supply of Ci for photosynthesis is less clear. The environmental factors that regulate calcification are still uncertain but appear to be related as much to the availability of nutrients as CO2. Transport of Ci to the intracellular site of calcification and removal of H+ from the coccolith vesicle appear to present few energetic or kinetic constraints. However, the large sustained transcellular fluxes of Ca2+ required for calcification probably occur via a pathway that does not involve diffusion across the cytoplasm.

Proteoparticles resemble TfR exosomes that are expelled by sheep reticulocytes upon maturation. The structure of proteoparticles in vitro is thus interpreted as being the result of the TfR's strong self-association potential, which might facilitate the endosomal sequestration of the TfR away from other membrane proteins and its subsequent return to the cell surface within tubular structures. The stalk is assumed to facilitate the tight packing of receptor molecules in coated pits and recycling tubuli.