Association of the alkaline phosphatase of rabbit polymorphonuclear leukocytes with the membrane of the specific granules.
The localization of alkaline phosphatase in the specific granules of rabbit polymorphonuclear leukocytes was investigated. The results obtained suggest very strongly that alkaline phosphatase is a component of the granule membrane. The enzyme remains attached to the membrane upon disruption of the granules by the use of detergents or by hypotonic shock and subsequent extraction with sodium sulfate, and can be isolated together with fragments of the granule membrane by isopycnic equilibration. Treatment of the granules with high amounts of Triton-X-100, sodium deoxycholate, or hexadecyltrimethylammonium bromide releases the enzyme in soluble form. In polymorphonuclear leukocyte homogenates, lysis of the granules is needed in order to render alkaline phosphatase fully accessible to substrates. This suggests that the catalytic site of the enzyme is exposed at the inner face of the granule membrane.
D000469:Alkaline Phosphatase / Q000097:blood*; D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000201:enzymology; D002499:Centrifugation, Density Gradient; D003594:Cytoplasmic Granules / Q000201:enzymology; D003840:Deoxycholic Acid / Q000494:pharmacology; D003902:Detergents / Q000494:pharmacology; D004072:Digitonin / Q000494:pharmacology; D007038:Hypotonic Solutions; D007962:Leukocytes / Q000201:enzymology*; D008854:Microscopy, Electron; D009113:Muramidase / Q000032:analysis; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology; D011817:Rabbits; D012964:Sodium / Q000494:pharmacology; D012965:Sodium Chloride / Q000494:pharmacology; D013431:Sulfates / Q000494:pharmacology; D013501:Surface-Active Agents / Q000494:pharmacology
D003902:Detergents; D007038:Hypotonic Solutions; D000644:Quaternary Ammonium Compounds; D013431:Sulfates; D013501:Surface-Active Agents; D003840:Deoxycholic Acid; D012965:Sodium Chloride; D012964:Sodium; D000469:Alkaline Phosphatase; D009113:Muramidase; D004072:Digitonin