The neutrophil glycoprotein Mo1 is an integral membrane protein of plasma membranes and specific granules.
The glycoprotein Mo1 has previously been demonstrated to be on the cell surface and in the specific granule fraction of neutrophils and to be translocated to the cell surface during degranulation. It is not known, however, whether Mo1 is an integral membrane protein or a soluble, intragranular constituent loosely associated with the specific granule membrane. Purified neutrophils were disrupted by nitrogen cavitation and separated on Percoll density gradients into four fractions enriched for azurophilic granules, specific granules, plasma membrane, and cytosol, respectively. The glycoproteins in these fractions were labeled with 3H-borohydride reduction, extracted with Triton X-114, and immunoprecipitated with 60.3, an anti-Mo1 monoclonal antibody Mo1 was detected only in the specific granule and plasma membrane fractions and partitioned exclusively into the detergent-rich fraction consistent with Mo1 being an integral membrane protein. In addition, treatment of specific granule membranes with a high salt, high urea buffer to remove absorbed or peripheral proteins failed to dissociate Mo1. These data support the hypothesis that Mo1 is an integral membrane protein of plasma and specific granule membranes in human neutrophils.
Stevenson
K B
KB
Department of Medicine, Veterans Administration Medical Center, Iowa City, IA.
Nauseef
W M
WM
Clark
R A
RA
D000911:Antibodies, Monoclonal / Q000276:immunology; D001798:Blood Proteins / Q000032:analysis* / Q000276:immunology; D003594:Cytoplasmic Granules / Q000032:analysis; D006801:Humans; D007425:Intracellular Membranes / Q000032:analysis; D016177:Macrophage-1 Antigen; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000276:immunology; D008565:Membrane Proteins / Q000032:analysis* / Q000276:immunology; D009504:Neutrophils / Q000032:analysis*
D000911:Antibodies, Monoclonal; D001798:Blood Proteins; D016177:Macrophage-1 Antigen; D008562:Membrane Glycoproteins; D008565:Membrane Proteins