Monoclonal antibodies to type IV collagenase recognize a protein with limited sequence homology to interstitial collagenase and stromelysin.
Type IV collagenase is a metalloproteinase associated with metastatic tumor cells. It specifically cleaves the triple helical basement membrane (type IV) collagen molecule at a single site. Monoclonal antibodies which block the activity of the human type IV collagenase were developed and used to purify this antigen. The purified type IV collagenase was partially sequenced following cyanogen bromide and trypsin cleavage. The amino acid sequence of the human type IV collagenase fragments revealed a region homologous to the human interstitial collagenase and stromelysin. However, several sequences in type IV collagenase were identified which are distinct from the latter. Polyclonal antibodies were raised against a synthetic peptide derived from such a sequence. Following affinity purification, the antibodies recognized the denatured human type IV collagenase in Western immunoblotting. These data indicate that type IV collagenase is a distinct member of a general family of metalloproteinases.
Höyhtyä
M
M
Biocenter, University of Oulu, Finland.
Turpeenniemi-Hujanen
T
T
Stetler-Stevenson
W
W
Krutzsch
H
H
Tryggvason
K
K
Liotta
L A
LA
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000941:Antigens / Q000276:immunology; D001485:Basement Membrane / Q000032:analysis; D001943:Breast Neoplasms / Q000201:enzymology; D003094:Collagen / Q000378:metabolism; D003488:Cyanogen Bromide; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007118:Immunoassay; D019278:Matrix Metalloproteinase 3; D008545:Melanoma / Q000201:enzymology; D008666:Metalloendopeptidases / Q000276:immunology*; D003012:Microbial Collagenase / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000201:enzymology*; D010446:Peptide Fragments; D012689:Sequence Homology, Nucleic Acid; D014357:Trypsin; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000941:Antigens; D010446:Peptide Fragments; D003094:Collagen; D014357:Trypsin; D008666:Metalloendopeptidases; D019278:Matrix Metalloproteinase 3; D003012:Microbial Collagenase; D003488:Cyanogen Bromide