Isolation and Characterization of Plasma Membranes from Transplantable Human Astrocytoma, Oat Cell Carcinoma, and Melanomas1
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چکیده
Purified plasma membranes were obtained from five transplantable human tumors, a Grade IV astrocytoma, an oat cell carcinoma, and three melanomas. Plasma membrane fractions were isolated from tumor homogenates by differential and discontinuous sucrose gradient centrifugation. Determination of enzyme activities indicated that the plasma membranes were enriched 10to 20-fold with respect to 5'-nucleotidase, nicotinamide adenine dinucleotide glycohydrolase, Mg2+-activated nucleoside triphosphatase, and sialic acid. Specific activities of nearly all the enzymes varied with the individual tumors, even among tumors of the same type, i.e., the melanomas. Electron micrographs of the plasma membrane fractions showed smooth single-membrane vesicles with slight contam ination by lysosomes. Therefore, these membranes are suitable for comparative biochemical studies and for the preparation of tumor-specific monoclonal antibodies. Plasma membranes from all five tumors contained very high Mg2+-adenosine triphosphatase (ATPase) activities. The Na+K+-ATPase was a minor component of the total ATPase of these membranes (<30%). The major component was an ATP ase exhibiting similar activity toward several nucleoside triphosphates. The activity of such a nucleoside triphosphatase has been correlated with tumorigenicity in cultured liver epithe lial cells. The nucleoside triphosphatase of the plasma mem branes of astrocytoma and oat cell carcinoma was stimulated from 50 to 100% by concanavalin A, whereas ATPase of the melanoma plasma membranes was not or only slightly stimu lated. The different response to concanavalin A could be due to differences in the ATPase molecules of the individual tumors or to the different environment of the ATPase.
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تاریخ انتشار 2006