Calpain secreted by activated human lymphoid cells degrades myelin.
Calpain secreted by lymphoid (MOLT-3, M.R.) or monocytic (U-937, THP-1) cell lines activated with PMA and A23187 degraded myelin antigens. The degradative effect of enzymes released in the extracellular medium was tested on purified myelin basic protein and rat central nervous system myelin in vitro. The extent of protein degradation was determined by SDS-PAGE and densitometric analysis. Various proteinase inhibitors were used to determine to what extent protein degradation was mediated by calpain and/or other enzymes. Lysosomal and serine proteinase inhibitors inhibited 20-40% of the myelin-degradative activity found in the incubation media of cell lines, whereas the calcium chelator (EGTA), the calpain-specific inhibitor (calpastatin), and a monoclonal antibody to m calpain blocked myelin degradation by 60-80%. Since breakdown products of MBP generated by calpain may include fragments with antigenic epitopes, this enzyme may play an important role in the initiation of immune-mediated demyelination.
Deshpande
R V
RV
Department of Neurology, Medical University of South Carolina, Charleston 29425, USA.
Goust
J M
JM
Hogan
E L
EL
Banik
N L
NL
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000494:pharmacology; D002154:Calpain / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology*; D002614:Chelating Agents / Q000494:pharmacology; D017077:Culture Media, Conditioned / Q000494:pharmacology; D003711:Demyelinating Diseases / Q000201:enzymology; D004533:Egtazic Acid / Q000494:pharmacology; D006801:Humans; D015459:Leukemia-Lymphoma, Adult T-Cell / Q000201:enzymology / Q000473:pathology; D016403:Lymphoma, Large B-Cell, Diffuse / Q000201:enzymology / Q000473:pathology; D009000:Monocytes / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D004676:Myelin Basic Protein / Q000378:metabolism*; D009186:Myelin Sheath / Q000187:drug effects*; D009363:Neoplasm Proteins / Q000378:metabolism / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D011817:Rabbits; D051381:Rats; D013601:T-Lymphocytes / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D002135:Calcium-Binding Proteins; D002614:Chelating Agents; D017077:Culture Media, Conditioned; D004676:Myelin Basic Protein; D009363:Neoplasm Proteins; D011480:Protease Inhibitors; D000001:Calcimycin; D004533:Egtazic Acid; C033668:calpastatin; D002154:Calpain; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium