Effects of monoclonal antibodies raised against the common acute lymphoblastic leukemia antigen on endopeptidase-24.11 activity.
The common acute lymphoblastic leukemia antigen (CALLA, CD10) has been identified as neutral endopeptidase-24.11 (NEP), a mammalian ectoenzyme involved in the inactivation of regulatory peptides, such as the enkephalins and atrial natriuretic peptide. Twenty monoclonal antibodies directed against the human antigen, were tested for their ability to inhibit the enzymatic activity of the human and rat peptidases expressed by cell lines. Six anti-CALLA antibodies were found to inhibit 50% or more of the hydrolysis of D-Ala2-leucine enkephalin by the neutral endopeptidase present on the human leukemic cell line Reh6 and, to a lesser extent, the hydrolysis of atrial natriuretic peptide. This may indicate that their binding may affect regions of the active site more important for the dipeptidylcarboxypeptidase activity of the enzyme. Only four antibodies cross-reacted with the peptidase from the rat epithelial cell line Rat2, as shown by membrane immunofluorescence, and these also partially inhibited enzyme activity. No antibody was able to inhibit completely the activity of the human and rat enzymes and all the active antibodies appeared to behave as non-competitive inhibitors of substrate cleavage. These monoclonal antibodies could be used in mapping studies of NEP.
Helene
A
A
Département de Chimie Organique, U 266 INSERM, UA 498 CNRS, UFR des Sciences Pharmaceutiques et Biologiques, Paris, France.
Milhiet
P E
PE
Haouas
H
H
Boucheix
C
C
Beaumont
A
A
Roques
B P
BP
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000943:Antigens, Differentiation / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000276:immunology*; D009320:Atrial Natriuretic Factor / Q000378:metabolism; D002460:Cell Line / Q000201:enzymology; D002462:Cell Membrane / Q000201:enzymology; D003429:Cross Reactions; D004743:Enkephalin, Leucine / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D015260:Neprilysin / Q000037:antagonists & inhibitors* / Q000276:immunology; D051381:Rats; D012689:Sequence Homology, Nucleic Acid
D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D000951:Antigens, Neoplasm; D004743:Enkephalin, Leucine; D009320:Atrial Natriuretic Factor; D015260:Neprilysin