Degradation of human erythrocyte surface components by human neutrophil elastase and cathepsin G: preferential digestion of glycophorins.
Human erythrocytes treated with purified human neutrophil elastase (HNE) or cathepsin G (CathG) were analysed by serological methods and by SDS-polyacrylamide gel electrophoresis followed by staining or immunoblotting with monoclonal antibodies. Both enzymes digested exhaustively glycophorins A, B and C, and HNE caused a partial digestion of band 3 protein. The degradation of other membrane proteins was not detectable by the methods used. Immunoblotting with the use of monoclonal antibodies against the defined epitopes of glycophorin A showed that HNE and CathG hydrolysed distinct peptide bonds in this antigen. The antibody PEP80, specific for the epitope in the cytoplasmic fragment of glycophorin A, gave patterns of bands which were characteristic for each of the two proteases. These bands could be distinctly identified in erythrocyte membrane samples containing only few percent of digested glycophorins. Therefore, the immunoblotting with this antibody may be useful as a sensitive assay for detecting the action of neutrophil proteases on red blood cells.
Bykowska
K
K
Institute of Haematology and Blood Transfusion, Warsaw, Poland.
Duk
M
M
Kusnierz-Alejska
G
G
Kopeć
M
M
Lisowska
E
E
D056649:Cathepsin G; D002403:Cathepsins / Q000494:pharmacology* / Q000502:physiology; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000187:drug effects*; D006021:Glycophorins / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D066298:In Vitro Techniques; D007519:Isoantigens / Q000187:drug effects; D019272:Leukocyte Elastase; D010196:Pancreatic Elastase / Q000494:pharmacology* / Q000502:physiology; D012697:Serine Endopeptidases
D006021:Glycophorins; D007519:Isoantigens; D002403:Cathepsins; D012697:Serine Endopeptidases; C534094:CTSG protein, human; D056649:Cathepsin G; D010196:Pancreatic Elastase; D019272:Leukocyte Elastase