Purification of ADAM 10 from bovine spleen as a TNFalpha convertase.
We have purified a protease with characteristics of TNFalpha convertase from bovine spleen membranes. Peptide sequencing of the purified protein identified it as ADAM 10 (Genbank accession no. Z21961). This metalloprotease cleaves a recombinant proTNFalpha substrate to mature TNFalpha, and can cleave a synthetic peptide substrate to yield the mature TNFalpha amino terminus in vitro. The enzyme is sensitive to a hydroxamate inhibitor of MMPs, but insensitive to phosphoramidon. In addition, cloned ADAM 10 mediates proTNFalpha processing in a processing-incompetent cell line.
Lunn
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Department of Immunology, Schering-Plough Research Institute, Kenilworth, NJ 07033, USA. charles.lunn@spcorp.com
Fan
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Dalie
B
B
Miller
K
K
Zavodny
P J
PJ
Narula
S K
SK
Lundell
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D
D051722:ADAM Proteins; D000072197:ADAM10 Protein; D000072198:ADAM17 Protein; D000595:Amino Acid Sequence; D053829:Amyloid Precursor Protein Secretases; D000818:Animals; D002417:Cattle; D002460:Cell Line; D003001:Cloning, Molecular; D004492:Edetic Acid / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006020:Glycopeptides / Q000494:pharmacology; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D008565:Membrane Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D008666:Metalloendopeptidases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D013154:Spleen / Q000201:enzymology*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D004791:Enzyme Inhibitors; D006020:Glycopeptides; D006877:Hydroxamic Acids; D008565:Membrane Proteins; D011498:Protein Precursors; D011994:Recombinant Proteins; C103777:SCH 43534; D014409:Tumor Necrosis Factor-alpha; C061153:tumor necrosis factor precursor; D004492:Edetic Acid; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D008666:Metalloendopeptidases; C103778:ADAM 10 protein, Bos taurus; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human; D000072198:ADAM17 Protein; C008890:phosphoramidon