Partial purification and characterization of neutral proteases in lymph nodes of rats with experimental allergic encephalomyelitis.
Two kinds of neutral protease activities in lymph nodes from Lewis rats with acute experimental allergic encephalomyelits (EAE) have been separated and partially purified and characterized. A soluble enzyme preparation enriched by gel filtration and ion-exchange chromatography hydrolyzes myelin basic protein, polylysine, and other basic proteins with an optimum pH at 6.0-6.5. It is inhibited by p-chloromercuribenzoate, and thus appears to be a mixture of thiol proteases. Another fraction containing proteolytic enzyme activity is strongly bound to the insoluble lymph node residue, and it also hydrolyzes myelin basic protein and histone, but not polylysine. It has a pH optimum above 7.5, is inhibited by phenylmethylsulfonyl fluoride, thus resembling elastase, but does not hydrolyze elastin-Congo red. The insoluble enzyme preparation hydrolyzes basic protein to 4-5 peptides in a pattern on polyacrylamide gels resembling that of the hydrolysis of basis protein by whole lymphocytes; the soluble enzyme mixture produces small fragments not retained on gels. Lymphocytes are a major component of the cells infiltrating the nervous system in experimental allergic encephalomyelitis, and neutral proteases contained in these cells may contribute to the degradation of myelin, especially of the basic protein.
D000818:Animals; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000201:enzymology*; D005260:Female; D006657:Histones / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008198:Lymph Nodes / Q000201:enzymology*; D008297:Male; D004676:Myelin Basic Protein / Q000378:metabolism; D010447:Peptide Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D011107:Polylysine / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology; D051381:Rats; D013379:Substrate Specificity
D006657:Histones; D004676:Myelin Basic Protein; D011480:Protease Inhibitors; D011107:Polylysine; D010447:Peptide Hydrolases