title: Use of GFP as a reporter for the analysis of sequence-specific proteases.
abstract: This unit describes a rapid fluorescent assay for sequence-specific proteases. A recombinant His-tagged substrate-GFP fusion protein containing the sequence-specific protease-recognition sequence is used as substrate. Batch metal-chelate chromatography separates uncleaved substrate-GFP fusion protein from GFP released by proteolysis and proteolytic activity is determined by measuring the fluorescence of GFP remaining in solution.
authors.affiliation: University of Nebraska, Lincoln, Nebraska.
authors.forename: Gautam
authors.initials: G
authors.lastname: Sarath
authors.affiliation: University of Memphis, Memphis, Tennessee.
authors.forename: Steven D
authors.initials: SD
authors.lastname: Schwartzbach
mesh_terms: D000595:Amino Acid Sequence; D005453:Fluorescence; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D010447:Peptide Hydrolases / Q000032:analysis*; D011993:Recombinant Fusion Proteins; D013379:Substrate Specificity
chemical_list: D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D010447:Peptide Hydrolases