[A method of screening artificial substrates for proteolytic enzymes].
A method of screening of proteolytic enzyme's substrates is proposed. An equimolar mixture of substrates consisting of peptide and easily detectable chromophore moieties (all chromophores in the mixture must be different) is subjected to enzymatic treatment. The cleaved chromophore groups, which are products of the substrate proteolysis, are quantitatively determined by chromatography. The Kcat/Km ratio is greater for substrates with higher initial rate accumulation of proteolysis products. The method is illustrated by screening of peptide derivatives of aminonaphtalene sulphonamides for trypsin assay. Proteolysis products are determined by HPLC with absorption detection or by TLC with fluorescence detection.
Nedospasov
A A
AA
Potaman
V N
VN
Rodina
E V
EV
D055598:Chemical Phenomena; D002621:Chemistry; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D006868:Hydrolysis; D010447:Peptide Hydrolases / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D013379:Substrate Specificity; D014357:Trypsin / Q000032:analysis
D010447:Peptide Hydrolases; D014357:Trypsin
Sposob skrininga iskusstvennykh substratov proteoliticheskikh fermentov.