Purification of endopeptidase-24.11 ('enkephalinase') from pig brain by immunoadsorbent chromatography.
Membrane preparations from striatum of pig brain contain endopeptidase activity towards iodoinsulin B-chain. Only 50% of the hydrolysis of insulin B-chain is inhibitable by phosphoramidon, and DEAE-cellulose chromatography can resolve the phosphoramidon-sensitive and -insensitive activities. The former activity (now designated 'endopeptidase-24.11') is responsible for hydrolysis of [D-Ala2,Leu5]enkephalin and is identical with an enzyme in brain that has previously been referred to as 'enkephalinase'. Pig striatal endopeptidase-24.11 has now been purified to homogeneity in a single step by immunoadsorbent chromatography using a monoclonal antibody. The overall purification was 23 000-fold, with a yield of 30%. The brain enzyme appears to be identical with kidney endopeptidase-24.11 in amino acid composition as well as by immunological and kinetic criteria. However, it differs slightly in apparent subunit size (Mr = 87 000), attributable to differences in glycosylation.
D000818:Animals; D000911:Antibodies, Monoclonal; D001921:Brain / Q000201:enzymology*; D002241:Carbohydrates / Q000032:analysis; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000276:immunology / Q000302:isolation & purification*; D006020:Glycopeptides / Q000494:pharmacology; D005779:Immunodiffusion; D007163:Immunosorbent Techniques; D007700:Kinetics; D008970:Molecular Weight; D015260:Neprilysin; D013552:Swine
D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D006020:Glycopeptides; D010450:Endopeptidases; D015260:Neprilysin; C008890:phosphoramidon