Detection of hydrolytic activity of trypsin with a fluorescence-chymotryptic peptide on a TLC plate.
To find a new trypsin-like enzyme, a simple assay method of the hydrolysis activity for trypsin has been found. We used 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) in the peptide labeling as a substrate for the trypsin-like peptidase in this study. The peptidase activity of trypsin was detected by using an AQC-chymotryptic peptide (AHP1) obtained from bovine hemoglobin. This showed that the substrate specificity of trypsin-like peptidase was distinguishable from that of the others by this procedure, and the method was used extensively in cases of various trypsin inhibitors with no significant interference from the concomitant.
Uchikoba
Tetsuya
T
Kagoshima University Musium, Kagoshima, Japan. uchik@sci.kagshima-u.ac.jp
Fukumoto
Shigeko
S
Itakura
Takao
T
Okubo
Michiko
M
Tomokiyo
Kazuhiko
K
Arima
Kazunari
K
Yonezawa
Hiroo
H
D000634:Aminoquinolines / Q000737:chemistry*; D000818:Animals; D002219:Carbamates / Q000737:chemistry*; D002417:Cattle; D002855:Chromatography, Thin Layer / Q000379:methods*; D005453:Fluorescence; D006454:Hemoglobins / Q000737:chemistry; D006868:Hydrolysis; D031426:Lycoris / Q000201:enzymology; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D010940:Plant Proteins / Q000032:analysis / Q000378:metabolism; D013379:Substrate Specificity; D014357:Trypsin / Q000032:analysis* / Q000378:metabolism*
C078178:6-aminoquinolyl-N-hydroxysuccinimidyl carbamate; D000634:Aminoquinolines; D002219:Carbamates; D006454:Hemoglobins; D010446:Peptide Fragments; D010940:Plant Proteins; C068347:Trichosanthes trypsin inhibitor; D014357:Trypsin