Clustering of integral membrane proteins of the human erythrocyte membrane stimulates autologous IgG binding, complement deposition, and phagocytosis.
Damaged or old erythrocytes are cleared rapidly from circulation. Because several common biochemical lesions can induce the clustering of integral membrane proteins, we have proposed that formation of microscopic protein aggregates in the membrane might constitute a cell surface marker that promotes removal of the defective/senescent cells. We demonstrate here that treatments that cluster integral membrane proteins in erythrocytes (1 mM ZnCl2, 1 mM acridine orange, and 0.35 microM melittin) induce autologous IgG binding, complement fixation, and phagocytosis by human monocytes in vitro. Removal of the clustering agents prior to incubation in autologous serum or cross-linking of cell surface proteins before addition of clustering agents prohibited the above response, while cross-linking after treatment with the clustering agents preserved the response even if the clustering agents were later removed. Furthermore, subsequent reversal of the chemical cross-link maintaining the clustered distribution also reversed the induction of IgG binding, complement deposition, and phagocytosis. Finally, by deleting or inactivating different steps in the phagocytosis pathway, the chronology of steps was shown to be: (i) integral protein clustering, (ii) IgG binding, (iii) complement deposition, and (iv) phagocytosis.
Turrini
F
F
Dipartimento di Genetica, Biologia e Chimica Medica, Universitá di Torino, Italy.
Arese
P
P
Yuan
J
J
Low
P S
PS
D000165:Acridine Orange / Q000494:pharmacology; D001457:Anion Exchange Protein 1, Erythrocyte / Q000187:drug effects / Q000302:isolation & purification / Q000502:physiology; D002478:Cells, Cultured; D002712:Chlorides / Q000494:pharmacology; D015932:Complement C3c / Q000378:metabolism; D003165:Complement System Proteins / Q000378:metabolism*; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000276:immunology / Q000502:physiology*; D004912:Erythrocytes / Q000187:drug effects / Q000502:physiology*; D005914:Globins / Q000187:drug effects / Q000302:isolation & purification / Q000502:physiology; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism*; D007700:Kinetics; D046911:Macromolecular Substances; D008555:Melitten / Q000494:pharmacology; D008565:Membrane Proteins / Q000097:blood* / Q000187:drug effects / Q000502:physiology; D009000:Monocytes / Q000502:physiology*; D010587:Phagocytosis; D013049:Spectrin / Q000187:drug effects / Q000302:isolation & purification / Q000502:physiology; D015032:Zinc / Q000494:pharmacology; D017967:Zinc Compounds
D001457:Anion Exchange Protein 1, Erythrocyte; D002712:Chlorides; D007074:Immunoglobulin G; D046911:Macromolecular Substances; D008565:Membrane Proteins; D017967:Zinc Compounds; D013049:Spectrin; D008555:Melitten; D015932:Complement C3c; C016837:zinc chloride; D005914:Globins; D003165:Complement System Proteins; D000165:Acridine Orange; D015032:Zinc