Purification and properties of 5'-nucleotidase from lymphocyte plasma membranes.
5'-Nucleotidase is purified from lymphocyte plasma membranes by two affinity chromatographies. The first one, on Lens culinaris lectin-Sepharose 4B yields a fraction of twelve lectin-binding glycoproteins (lectin-receptor fraction). The second one on 5'-AMP-Sepharose 4B leads to pure enzyme. This enzyme is a glycoprotein with a molecular weight of 130 000; it gives a single band in polyacrylamide/dodecylsulfate electrophoresis and displays a very high specific activity (2500-3000 mumol Pih-1mg-1). Some properties of purified 5'-nucleotidase are similar to those of membrane-bound enzyme: substrate specificity, temperature dependence, effects of ions and SH-blocking reagents. Others are completely different for the two systems and these differences result from an interaction between the enzyme molecule and other Lens culinaris lectin binding proteins.
D000818:Animals; D002413:Cations, Divalent / Q000494:pharmacology; D002462:Cell Membrane / Q000201:enzymology; D002846:Chromatography, Affinity; D006863:Hydrogen-Ion Concentration; D037102:Lectins; D008214:Lymphocytes / Q000201:enzymology*; D008970:Molecular Weight; D009708:Nucleotidases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D011952:Receptors, Concanavalin A; D013379:Substrate Specificity; D013552:Swine; D013696:Temperature
D002413:Cations, Divalent; D037102:Lectins; D011952:Receptors, Concanavalin A; D009708:Nucleotidases