title: Subcellular localization and properties of adenosine diphosphatase activity in human polymorphonuclear leukocytes.
abstract: Adenosine diphosphatase (ADPase) activities were studied in human polymorphonuclear leukocytes with a recently developed radio-assay. The neutrophils were homogenized in isotonic sucrose and subjected to analytical subcellular fractionation. The sucrose density gradient fractions were assayed for ADPase activity and for principal organelle marker enzymes. ADPase activity was distributed between the plasma membrane, specific granule and soluble fractions. The plasma membrane and specific granule activities had similar kinetic and inhibitor properties but the cytosolic enzyme was clearly different. Studies with the non-penetrating inhibitor diazotized sulphanilic acid and measurements of latent activity indicate that plasma membrane ADPase activity is located on the external aspect to the cell. Its possible role in inhibiting platelet aggregation is discussed. Neutrophils were isolated from control subjects, patients with chronic granulocytic leukaemia and patients in the third trimester of pregnancy. The specific activities (mU/mg protein) of ADPase activity, in contrast to those of alkaline phosphatase, were similar in all three groups. This result, together with fractionation experiments and inhibition studies strongly suggests that ADPase activity is not attributable to neutrophil alkaline phosphatase.
mesh_terms: D000469:Alkaline Phosphatase / Q000097:blood; D001081:Apyrase / Q000097:blood*; D002462:Cell Membrane / Q000201:enzymology; D002499:Centrifugation, Density Gradient; D055598:Chemical Phenomena; D002621:Chemistry; D003600:Cytosol / Q000201:enzymology; D005260:Female; D006801:Humans; D007700:Kinetics; D007951:Leukemia, Myeloid / Q000201:enzymology; D009504:Neutrophils / Q000201:enzymology*; D010744:Phosphoric Monoester Hydrolases / Q000097:blood*; D011247:Pregnancy; D013347:Subcellular Fractions / Q000201:enzymology
chemical_list: D000469:Alkaline Phosphatase; D010744:Phosphoric Monoester Hydrolases; D001081:Apyrase