Localization of long-chain fatty acids and unconventional sterols in spherulous cells of a marine sponge.

The first direct evidence is provided for the presence of unconventional lipids in a particular subcellular membrane system of a sponge. Spherulous cells were isolated from the variety of cell types present in the marine sponge Aplysina fistularis by density gradient centrifugation. Spherulous cell plasma membrane was subsequently isolated by cell rupture followed by differential centrifugation and sucrose, or Percoll, density gradient ultracentrifugation. Plasma membrane isolates were identified and assessed for purity using [3H]concanavalin A plasma membrane marker, sodium dodecyl sulfate polyacrylamide gel electrophoresis and ratios of protein, sterol and phosphate. Plasma membrane isolates could not be assessed for purity by traditional enzymatic means. Spherulous cell plasma membrane was found to contain unusual lipids, including long-chain (C24-C30) fatty acids (16.8-27.2%) and unconventional 26-alkylated sterols (66.4-72.6%), in addition to more conventional fatty acids and sterols. Spherulous cell intracellular membranes were also found to contain long-chain fatty acids and unconventional sterols, although the relative importance of these unusual lipids apparently varies between intracellular membranes, with some containing approximately 50% long-chain acids.