The glycosylphosphatidylinositol-anchored form and the transmembrane form of CD58 are released from the cell surface upon antibody binding.
The adhesion molecule CD58 is expressed on the cell surface in both a transmembrane form and a glycosylphosphatidylinositol (GPI)-anchored form. Here we report that CD58 is released from JY cells following cross-linking by immobilized anti-CD58 monoclonal antibodies. Antibodies to other cell surface proteins, as well as PMA and LPS, did not trigger CD58 release. The release resulted from membrane cleavage, since biotin-labeled CD58 was released from biotinylated cells, and down-modulation of CD58 surface expression accompanied accumulation of soluble CD58 IN culture media. We have previously reported the isolation of JY variant cells, which lack expression of GPI anchored proteins and thus express only the transmembrane form of CD58. Here we show that these variant cells release CD58 upon crosslinking, indicating that the transmembrane isoform is released, probably by proteolysis. Antibodies directed to the cytoplasmic domain of CD58, in contrast to antibodies against an extracellular epitope of CD58, did not react with released CD58, supporting a membrane cleavage mechanism. It is also shown that CD58, released from [3H]ethanolamine-labeled JY cells, contained ethanolamine. This result demonstrated that the GPI-anchored CD58 can be released in parallel to the transmembrane isoform and that this release does not result from proteolytic cleavage, since cleavage by a protease would have removed the ethanolamine. The present data suggest that the two isoforms of CD58 are released upon antibody binding and that their release is mediated by distinct mechanisms.
Itzhaky
D
D
Department of Human Microbiology, Sackler School of Medicine, Tel-Aviv University, Israel.
Raz
N
N
Hollander
N
N
D000937:Antigen-Antibody Reactions; D018968:CD58 Antigens / Q000276:immunology / Q000378:metabolism*; D002460:Cell Line; D017261:Glycosylphosphatidylinositols / Q000502:physiology*; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D008970:Molecular Weight
D018968:CD58 Antigens; D017261:Glycosylphosphatidylinositols; D008565:Membrane Proteins