Spontaneous and detergent-induced vesiculation of thymocyte plasma membranes.
An analog of lysophosphatidylcholine (1-dodecyl-propanediol-3-phosphocholine) which does not impair membrane-bound enzymes was used for the induction of shedding of membrane vesicles from intact calf thymocytes. Without liberation of intracellular enzymes such as lactate dehydrogenase (EC 1.1.1.27) the shedded membranes contained 15--25% of the total activity of the plasma membrane enzymes alkaline phosphatase (EC 3.1.3.1), nucleotide pyrophosphatase (EC 3.1.4.1) and gamma-glutamyl transferase (EC 2.3.2.2). Membrane-free supernatants only exhibited trace activities of these enzymes. Without further purification, the specific enzyme activities in shedded membranes were of the same order of magnitude as in purified plasma membranes prepared after nitrogen cavitation of thymocytes. Small amounts of membrane vesicles which showed a different composition could be removed without detergent. These membranes exhibited a 3-fold lower specific activity of the gamma-glutamyl transferase while that of the alkaline phosphatase and nucleotide pyrophosphatase was similar as in detergent induced membrane vesicles. Distinct differences also were found in the protein pattern. The content of total cholesterol and phospholipid in vesicles shed spontaneously or after detergent treatment was nearly identical, however, significant differences were found in the fatty acid composition of the main phospholipids. The content of polyunsaturated fatty acids (linoleic and arachidonic acid) increased in the order: spontaneously shedded membranes, detergent induced vesicles, conventional purified plasma membranes. These results are discussed in terms of the heterogeneous composition of areas of the thymocyte plasma membrane.
D000118:Acetylglucosaminidase / Q000032:analysis; D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure*; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D008563:Membrane Lipids / Q000032:analysis; D010744:Phosphoric Monoester Hydrolases / Q000032:analysis; D013950:Thymus Gland / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure*; D014166:Transferases / Q000032:analysis
D008563:Membrane Lipids; D007770:L-Lactate Dehydrogenase; D014166:Transferases; D010744:Phosphoric Monoester Hydrolases; D000118:Acetylglucosaminidase