[New fluorescent substrates for metalloendopeptidases with internal quenching of fluorescence].
p-Nitroanilides of antranyloyltripeptides of the general structure Abz-Ala-Ala-P'1-pNA (P'1 = Phe, Leu, Ile, Val) containing intramolecularly quenched fluorescent groups (Abz is a fluorogenic group and pNA is a quencher of fluorescence) were prepared by combination of chemical and enzymatic methods. Thermolysin and metalloproteinases from Legionella pneumophila and Thermoactinomyces species were shown to hydrolyse Ala-P'1 bond of the peptides with simultaneous 4-7 fold increase in fluorescence. Kinetic parameters for enzymatic hydrolysis of the substrates were determined. Metalloendopeptidases can be assayed in the presence of serine proteinases (of the subtilisin type) using Abz-Ala-Ala-Ile-pNA or Abz-Ala-Ala-Val-pNA.
I Iu
IIu
Filippova
E N
EN
Lysogorskaia
E S
ES
Oksenoĭt
E P
EP
Troshchenkova
V M
VM
Stepanov
D000190:Actinomyces / Q000201:enzymology; D005456:Fluorescent Dyes / Q000138:chemical synthesis*; D006868:Hydrolysis; D007700:Kinetics; D007875:Legionella / Q000201:enzymology; D008666:Metalloendopeptidases / Q000032:analysis*; D013379:Substrate Specificity
D005456:Fluorescent Dyes; D008666:Metalloendopeptidases
Novye fluorestsentnye substraty metalloéndopeptidaz s vnutrennim tusheniem fluorestsentsii.