Anti-peptide antibodies to cathepsins B, L and D and type IV collagenase. Specific recognition and inhibition of the enzymes.
Anti-peptide antibodies were raised against synthetic peptides selected from the sequences of human cathepsins B and L, porcine cathepsin D and human type IV collagenase. Sequences were selected from the active site clefts of the cathepsins in the expectation that these would elicit immunoinhibitory antibodies. In the case of type IV collagenase a sequence unique to this metalloproteinase subclass and suitable for immunoaffinity purification, was chosen. Antibodies against the chosen cathepsin B sequence were able to recognize the peptide but were apparently unable to recognise the whole enzyme. Antibodies against the chosen cathepsin L sequence were found to recognise and inhibit the native enzyme and were also able to discriminate between denatured cathepsins L and B on Western blots. Antibodies against the chosen cathepsin D sequence recognised native cathepsin D in a competition ELISA, but did not inhibit the enzyme. Native type IV collagenase was purified from human leukocytes by immuno-affinity purification with the corresponding anti-peptide antibodies.
Coetzer
T H
TH
Department of Biochemistry, University of Natal, Pietermaritzburg, Republic of South Africa.
Elliott
E
E
Fortgens
P H
PH
Pike
R N
RN
Dennison
C
C
D000595:Amino Acid Sequence; D000917:Antibody Formation; D000918:Antibody Specificity; D015153:Blotting, Western; D002401:Cathepsin B / Q000276:immunology; D002402:Cathepsin D / Q000276:immunology; D056668:Cathepsin L; D002403:Cathepsins / Q000276:immunology*; D003546:Cysteine Endopeptidases; D010450:Endopeptidases; D004792:Enzyme Precursors / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D020780:Matrix Metalloproteinase 9; D003012:Microbial Collagenase / Q000276:immunology*; D008969:Molecular Sequence Data
D004792:Enzyme Precursors; D002403:Cathepsins; D010450:Endopeptidases; D003546:Cysteine Endopeptidases; D002401:Cathepsin B; C534281:CTSL protein, human; D056668:Cathepsin L; D002402:Cathepsin D; D003012:Microbial Collagenase; D020780:Matrix Metalloproteinase 9