Detection of protease activity with a fluorescence-labelled peptide substrate on a TLC plate.
A small amount of peptidase activity could be detected using an amine derivatizing reagent, fluorescein isothiocyanate (FITC), which has been used to produce a fluorogenic peptide. The substrate produced, FITC-peptide, gave a clear spot on a silica gel sheet upon exposure to UV light. The peptidase activity of angiotensin-converting enzyme (ACE), trypsin, chymotrypsin, cucumisin, and that of some plant tissues were detected by using a fluorogenic angiotensin I. This showed that the substrate specificity of proteolytic enzymes can be distinguished from the others by this procedure.
Department of Chemistry, Faculty of Science, Kagoshima University, Korimoto, Japan. uchik@sci.kagoshima-u.ac.jp
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T
Uchikoba
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H
Yonezawa
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K
Arima
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M
Kaneda
D000803:Angiotensin I; D000818:Animals; D002855:Chromatography, Thin Layer; D002918:Chymotrypsin / Q000032:analysis; D010450:Endopeptidases / Q000032:analysis*; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006801:Humans; D007703:Peptidyl-Dipeptidase A / Q000032:analysis; D010944:Plants / Q000201:enzymology; D012697:Serine Endopeptidases / Q000032:analysis; D014357:Trypsin / Q000032:analysis
D005456:Fluorescent Dyes; D000803:Angiotensin I; D010450:Endopeptidases; D007703:Peptidyl-Dipeptidase A; D012697:Serine Endopeptidases; D002918:Chymotrypsin; C041541:cucumisin; D014357:Trypsin; D016650:Fluorescein-5-isothiocyanate