CD63 associates with CD11/CD18 in large detergent‐resistant complexes after translocation to the cell surface in human neutrophils 1

• Published in: FEBS letters Vol. 469; no. 1; pp. 52 - 56
• Main Authors: Skubitz, Keith M., Campbell, Kenneth D., Skubitz, Amy P.N.
• Format: Journal Article
• Language: English
• Published: 03.03.2000
• Subjects: Brij solubilization buffer, 20 mM Tris–HCl, pH 8.2, 150 mM NaCl, 1 mM PMSF, 2 mM MgCl2, 0.02% NaN3 and 1.0% Brij 58; Brij wash buffer, 20 mM Tris–HCl, pH 8.2, 150 mM NaCl, 1 mg/ml BSA, 0.5% Brij 58, 2 mM MgCl2, 0.125 mg/ml gelatin, 1 mM PMSF and 0.02% NaN3; CD63; DFP, diisopropylfluorophosphate; GPI, glycosyl-phosphatidylinositol; Granulocyte; Inflammation; Integrin; Membrane domain; Neutrophil activation; NMS, normal mouse serum; PBS, phosphate buffered saline, pH 7.4; SDS–PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(00)01240-0

CD63 antibody binding to the neutrophil surface triggers a transient activation signal that regulates the adhesive activity and surface expression of CD11/CD18. Gel permeation chromatography demonstrated that all of the cell surface CD11/CD18 associated with CD63 eluted in the void volume, indicating that they were present in large detergent‐resistant complexes. In contrast, the majority of the total cellular CD63, CD11 and CD18, which are largely intracellular, was not present in complexes. The data suggest that intracellular CD11, CD18 and CD63 are not in detergent‐resistant complexes, but enter such complexes following translocation to the cell surface.