Ecto-enzymes of mammary gland and its tumors. Lectin inhibition of 5'-nucleotidase of the 13762 rat mammary ascites carcinoma.
Lectin inactivation of the cell surface enzyme 5'-nucleotidase has been studied in intact 13762 mammary ascites cells, cell membrane envelopes, membrane vesicles, and solubilized membranes as a means of understanding the effects of lectins on biochemical processes and the behavior of the enzyme in the membranes. The properties of the enzyme are essentially the same in the intact cells and in membrane envelopes prepared after zinc treatment of the cells. The enzyme has a Km of 25 muM and is inhibited by concanavalin A (Con A) by an apparent noncompetitive process, which is reversed by alpha-methylmannoside. Half-maximal inhibition requires about 50 mug of Con A per mg of membrane protein. The inhibition for the intact cells and membrane envelopes does not exhibit significant cooperativity. Membrane vesicles, prepared by EDTA extraction, and solubilized membranes show differences in the behavior of the nucleotidase toward Con A even though the Km of the enzyme (25 muM) and inactivation (noncompetitive) are essentially unchanged. The Hill coefficient for the inactivation process in solubilized membranes and membrane vesicles is shifted to near 2, indicating significant cooperativity in the Con A-enzyme interaction. It is suggested that this change in cooperativity may reflect a different mode of interaction with the membrane and increased enzyme mobility, although an alteration in enzyme subunit interactions cannot be ruled out at present.
D000818:Animals; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology; D003208:Concanavalin A / Q000494:pharmacology*; D003840:Deoxycholic Acid; D005260:Female; D007700:Kinetics; D037102:Lectins / Q000494:pharmacology*; D008325:Mammary Neoplasms, Experimental / Q000201:enzymology*; D008766:Methylmannosides / Q000494:pharmacology; D009708:Nucleotidases / Q000037:antagonists & inhibitors*; D011092:Polyethylene Glycols; D051381:Rats
D037102:Lectins; D008766:Methylmannosides; D003840:Deoxycholic Acid; D003208:Concanavalin A; D011092:Polyethylene Glycols; D009708:Nucleotidases