CD23 (FcepsilonRII) release from cell membranes is mediated by a membrane-bound metalloprotease.
CD23 (low-affinity IgE receptor, FcepsilonRII) is expressed as a Type II extracellular protein on a variety of cells such as B-cells, monocytes and macrophages and is cleaved from the cell surface to generate several distinct fragments. The expression of CD23 on the cell surface as well as the generation of soluble fragments of CD23 has been shown to be involved in the regulation of IgE synthesis. Here we report that the release of CD23 from the cell surface is mediated by a metalloprotease. An assay utilizing purified CD23 and an neo-epitope antibody specific for one of the known cleavage products is described and used to demonstrate unambiguously the cleavage of CD23 by a distinct protease. Characterization of the mechanism of CD23 processing shows that the protease exists as an integral membrane protein with a functional molecular mass of approx. 63 kDa as determined by gel-filtration chromatography. The CD23-cleaving activity found in enriched plasma membranes from RPMI 8866 cells is inhibited by the metalloprotease inhibitors 1, 10-phenanthroline and imidazole and by the matrix metalloprotease inhibitor batimastat, but not by inhibitors of cysteine proteases, serine proteases or acid proteases. The same or a similar activity that cleaves CD23 to the known 33 kDa fragment and is inhibited by batimastat is present in diverse cell types such as unstimulated fibroblasts and monocytic cell lines not expressing CD23, as well as in the Epstein-Barr virus-transformed B-cell line, RPMI 8866, which constitutively expresses CD23.
Marolewski
A E
AE
SmithKline Beecham Pharmaceuticals, Upper Merion, 709 Swedeland Road, King of Prussia, PA 19406, USA.
Buckle
D R
DR
Christie
G
G
Earnshaw
D L
DL
Flamberg
P L
PL
Marshall
L A
LA
Smith
D G
DG
Mayer
R J
RJ
D000818:Animals; D001402:B-Lymphocytes / Q000201:enzymology / Q000648:ultrastructure; D002462:Cell Membrane / Q000201:enzymology / Q000378:metabolism; D002478:Cells, Cultured; D002850:Chromatography, Gel; D006224:Cricetinae; D005347:Fibroblasts / Q000201:enzymology / Q000648:ultrastructure; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008970:Molecular Weight; D009000:Monocytes / Q000201:enzymology / Q000648:ultrastructure; D017455:Receptors, IgE / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012995:Solubility; D013601:T-Lymphocytes / Q000201:enzymology / Q000648:ultrastructure
D017455:Receptors, IgE; D008666:Metalloendopeptidases