Rabbit small intestinal brush border membrane preparation and lipid composition.

1. Rabbit intestinal brush border membrane vesicles were prepared either from frozen or fresh tissue and the lipid composition was analysed. 2. Lipids extracted from membranes prepared by the Ca2+-precipitation method (Schmitz, J., Preiser, H., Maestracci, D., Ghosh, B.K., Cerda, J.J. and Crane, R.K. (1973) Biochim. Biophys. Acta 323, 98--112; Kessler, M., Acuto, O., Storelli, C., Murer, H., Müller, M. and Semenza, G. (1978) Biochim. Biophys. Acta 506, 136--154) had exceptionally high levels of lysophospholipids and free fatty acids. An intrinsic, Ca2+-activated phospholipase A is responsible for the lipid decomposition. 3. It was necessary to modify the preparation of brush border membranes. Essentially, EGTA is used to keep the free Ca2+ concentration low so that phospholipases are inactivated, and Mg2+ instead of Ca2+ is employed to aggregate selectively contaminating membranes. The modified procedure gives membranes suitable for lipid analysis. 4. The molar ratio of neutral, phospholipid and glycolipid is about 1 : 1 : 1. The major neutral lipids are free cholesterol and fatty acids. The major phospholipids are phosphocholine-containing (approx. 45%) and phosphatidyl-ethanolamine (approx. 40%). The remainder (15--20%) is made up ppf acidic phospholipids, mainly phosphatidylserine and phosphatidylinositol. The major glycolipid is ceramide monohexodise. 5. The major fatty acids of total lipids and phospholipids are palmitic, stearic, oleic and linoleic acids. Individual phospholipids are characterised by distinct fatty acid compositions and differ markedly in the ratio of unsaturated-to-saturated fatty acid. X