Studies on the kinetic properties and subcellular localization of adenosine diphosphatase activity in human peripheral blood lymphocytes.
Using a recently developed radioassay, the conditions for measuring adenosine diphosphatase (ADPase) activity in human lymphocytes were optimized. Kinetic studies showed that the activity was optimal at pH 8.0 and required 0.2 mM MgCl2. The Km of the enzyme for ADP was 0.03 mM. Analytical subcellular fractionation showed that the ADPase activity was distributed between the plasma membrane and mitochondria. Studies with the non-penetrating inhibitor, diazotized sulphanilic acid, indicate that plasma membrane ADPase activity is located on the external aspect of the cell. Polymorphonuclear leucocytes were found to have about three times the ADPase activity. Lymphocytes, therefore, have the ability to generate AMP on their surface which can be further metabolized to adenosine by ecto-5' nucleotidase. A role for this system is discussed.
D001081:Apyrase / Q000097:blood*; D002462:Cell Membrane / Q000201:enzymology; D002500:Centrifugation, Isopycnic; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008214:Lymphocytes / Q000201:enzymology*; D008928:Mitochondria / Q000201:enzymology; D010744:Phosphoric Monoester Hydrolases / Q000097:blood*
D010744:Phosphoric Monoester Hydrolases; D001081:Apyrase