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.
Uchikoba
T
T
Department of Chemistry, Faculty of Science, Kagoshima University, Korimoto, Japan. uchik@sci.kagoshima-u.ac.jp
Yonezawa
H
H
Arima
K
K
Kaneda
M
M
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