Neutrophil-specific antigen NB1 is anchored via a glycosyl-phosphatidylinositol linkage.
Neutrophil-specific alloantibodies and the antigens they recognize are important in clinical medicine but little is known about the structure of these antigens. Alloimmunization to the antigen NB1 is a clinically important cause of neonatal neutropenia and leukocyte-mediated transfusion reactions. A novel mechanism of protein attachment to cell membranes involving the covalent linkage of the protein through an oligosaccharide to phosphatidylinositol has recently been defined. Many proteins which are anchored to the cell membrane by this mechanism can be released by treatment with phosphatidylinositol-specific phospholipase C (PI-PLC). The 58-64-kDa human neutrophil surface protein which contains the NB1 antigen was labeled with 125I by using lactoperoxidase and examined for PI-PLC sensitivity. The 58-64-kDa protein was specifically released from the cell by treatment with PI-PLC, and the mobility of the protein under non-denaturing conditions using non-ionic detergent was increased by treatment with PI-PLC. Surface expression of the NB1 antigen was slightly up-regulated by treatment with the chemotactic peptide f-met-leu-phe. Removal of N-linked carbohydrates with endoglycosidase-F decreased the apparent molecular weight of the protein to approximately 45-kDa. The data suggest that most of the 58-64-kDa protein bearing the neutrophil-specific antigen NB1 is anchored to the membrane through a glycosyl-phosphatidylinositol linkage.
Skubitz
K M
KM
Department of Medicine, University of Minnesota Medical School, Minneapolis.
Stroncek
D F
DF
Sun
B
B
D000954:Antigens, Surface / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D058851:GPI-Linked Proteins; D006017:Glycolipids / Q000032:analysis*; D006026:Glycoside Hydrolases; D017261:Glycosylphosphatidylinositols; D006801:Humans; D007519:Isoantigens / Q000276:immunology; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008562:Membrane Glycoproteins / Q000032:analysis*; D009504:Neutrophils / Q000276:immunology*; D010716:Phosphatidylinositols / Q000032:analysis*; D011956:Receptors, Cell Surface; D012016:Reference Values
D000954:Antigens, Surface; C407242:CD177 protein, human; D058851:GPI-Linked Proteins; D006017:Glycolipids; D017261:Glycosylphosphatidylinositols; D007519:Isoantigens; D008562:Membrane Glycoproteins; D010716:Phosphatidylinositols; D011956:Receptors, Cell Surface; D006026:Glycoside Hydrolases; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase