title: Detection and analysis of neutral endopeptidase from tissues with substrate gel electrophoresis.
abstract: Neutral endopeptidase from human or bovine tissues retains enzymatic activity following electrophoresis and immobilization in polyacrylamide gels. Infiltration of the gel with a fluorogenic substrate permits identification of the active enzyme by fluorescence associated with a distinct protein band. This technique both separates and identifies the enzymatically active species from a crude cell membrane fraction or from partially purified extracts that contain contaminating proteins. Enzymatic activity is quantitated by photographing the fluorescent bands and scanning the negatives with a laser densitometer. Because as little as 25 ng of enzyme can be detected by this method, it could be used where the amount of material is limited.
authors.affiliation: Department of Biochemistry, University of Texas Health Center, Tyler 75710.
authors.forename: J
authors.initials: J
authors.lastname: Sullivan
authors.forename: A R
authors.initials: AR
authors.lastname: Johnson
mesh_terms: D000818:Animals; D002417:Cattle; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004800:Enzymes, Immobilized; D005333:Fetus; D006031:Glycosylation; D006801:Humans; D007668:Kidney / Q000201:enzymology*; D008022:Ligaments / Q000201:enzymology*; D008970:Molecular Weight; D015260:Neprilysin / Q000302:isolation & purification* / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D004800:Enzymes, Immobilized; D015260:Neprilysin