A sensitive colorimetric assay for human urinary kallikrein.
L-Prolyl-L-phenylalanyl-L-arginine-alpha-naphthylester (Pro-Phe-Arg-NE) was synthesized as a new substrate for use in the assay of kallikreins. An assay was developed based on the colorimetric determination of alpha-naphthol released by the enzyme reaction. With Pro-Phe-Arg--NE as substrate, the minimum detectable concentration of human urinary kallikrein, was about 0.001 KU. Thus use of Pro-Phe-Arg-NE provides a highly sensitive method for detection of human urinary kallikrein. Kallikrein could be determined with a 25-microliter sample of human urine. Zymograms of human urinary kallikrein were prepared using Pro-Phe-Arg-NE as substrate. Six bands were separated by polyacrylamide disc gel isoelectrophoresis.
D003124:Colorimetry / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006868:Hydrolysis; D007525:Isoelectric Focusing; D007610:Kallikreins / Q000652:urine*; D010447:Peptide Hydrolases; D010455:Peptides / Q000138:chemical synthesis; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D013379:Substrate Specificity
D010455:Peptides; D012967:Sodium Dodecyl Sulfate; D010447:Peptide Hydrolases; D007610:Kallikreins