A zinc metalloproteinase is responsible for the release of CD30 on human tumor cell lines.
The activation marker CD30 is expressed on the cell surface of the malignant cells in Hodgkin's disease and a few non-Hodgkin lymphomas. We have analyzed the regulation of membrane-bound CD30 and found that the binding of a variety of anti-CD30 antibodies induced down-regulation of CD30 on cell lines. In addition, such down-modulation was also observed after treatment of the cell surface proteins with the sulfhydryl reagent iodoacetamide or after stimulation of the second messenger pathway with phorbol ester or calcium ionophore. This modulation was abolished at 4 degrees C and strongly inhibited by chelators like EDTA or 1,10-phenanthroline, whereas EGTA, a selective inhibitor of Ca(2+)-dependent proteinases and other inhibitors of serine, thiol and acid proteinases, showed no effect. The down-modulation was strengthened by Zn2+ or Cd2+, but not by other divalent cations such as Fe2+, Mn2+, Mg2+, Ca2+ or Co2+, thus indicating the involvement of a zinc metalloproteinase in CD30 modulation which can be activated by protein kinase C and by alkylation of sulfhydryl groups. Pulse-chase experiments, analysis of the CD30 glycosylation and specific measurement of the 90-kDa soluble form of CD30 (sCD30) with a sandwich radioimmunoassay revealed that CD30 down-modulation results from enhanced release of 90-kDa sCD30 by the site-specific cleavage of CD30 accomplished by a zinc metalloproteinase. This release occurs at the cell membrane without prior endocytosis.
Hansen
H P
HP
Department of Biochemistry, University of Kiel, Germany.
Kisseleva
T
T
Kobarg
J
J
Horn-Lohrens
O
O
Havsteen
B
B
Lemke
H
H
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D000001:Calcimycin / Q000494:pharmacology; D015536:Down-Regulation / Q000187:drug effects; D006031:Glycosylation; D006689:Hodgkin Disease / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D007460:Iodoacetamide / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D017730:Ki-1 Antigen / Q000378:metabolism*; D007700:Kinetics; D008228:Lymphoma, Non-Hodgkin / Q000201:enzymology / Q000378:metabolism*; D008666:Metalloendopeptidases / Q000378:metabolism*; D012995:Solubility; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D007476:Ionophores; D017730:Ki-1 Antigen; D013439:Sulfhydryl Reagents; D000001:Calcimycin; D008666:Metalloendopeptidases; D013755:Tetradecanoylphorbol Acetate; D007460:Iodoacetamide