Use of GFP as a reporter for the analysis of sequence-specific proteases.
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.
Sarath
Gautam
G
University of Nebraska, Lincoln, Nebraska.
Schwartzbach
Steven D
SD
University of Memphis, Memphis, Tennessee.
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
D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D010447:Peptide Hydrolases