Shedding and enrichment of the glycolipid-anchored complement lysis inhibitor protectin (CD59) into milk fat globules.
Protectin (CD59) is a glycolipid-anchored inhibitor of the membrane attack complex (MAC) of human complement (C) that protects blood cells, endothelial cells and various epithelial cells from C-mediated lysis. Because of its activities protectin is a candidate molecule for use in the treatment of paroxysmal nocturnal haemoglobinuria or conditions where MAC causes tissue damage. Soluble, phospholipid-free forms of protectin have been isolated from human urine and produced in recombinant form, but they have only a relatively weak C lysis-inhibiting activity. In the present study we have looked for functionally active protectin in human breast milk. Milk is rich in fat droplets, milk fat globules (MFG), that are enveloped in a plasma membrane derived from secretory cells of the mammary gland. The membranes of MFG contain a variety of glycoproteins expressed by the mammary epithelial cells. Both immunofluorescence and immunoblotting analysis demonstrated that protectin was strongly expressed on human MFG. In sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, MFG protectin (CD59M) appeared as distinct bands with apparent molecular weights of 19,000-23,000 MW, similar to protectin extracted from MCF7 breast carcinoma cells. CD59M in breast milk was functionally active and had a glycophospholipid anchor, as judged by its ability to incorporate into guinea-pig erythrocytes and inhibit their lysis by human complement. These results indicate that functionally active protectin becomes enriched in MFG and imply that secretion of glycophospholipid-anchored molecules, e.g. into cow milk and colostrum, could be exploited as a means of producing bioactive molecules that need to be targeted into cell membranes.
Hakulinen
J
J
Department of Bacteriology and Immunology, University of Helsinki, Finland.
Meri
S
S
D018957:CD59 Antigens / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D003169:Complement Inactivator Proteins / Q000737:chemistry / Q000378:metabolism*; D015938:Complement Membrane Attack Complex / Q000378:metabolism; D003602:Cytotoxicity, Immunologic; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D008856:Microscopy, Fluorescence; D008895:Milk, Human / Q000276:immunology* / Q000378:metabolism; D008970:Molecular Weight
D018957:CD59 Antigens; D003169:Complement Inactivator Proteins; D015938:Complement Membrane Attack Complex