73536-37-5Relevant academic research and scientific papers
Highly specific substrates of proteinase 3 containing 3-(2-Benzoxazol-5-yl) - l -alanine and their application for detection of this enzyme in human serum
Wysocka, Magdalena,Lesner, Adam,Guzow, Katarzyna,Kulczycka, Julia,Legowska, Anna,Wiczk, Wieslaw,Rolka, Krzysztof
, p. 3883 - 3889 (2010)
A set of benzoxazolyl-l-alanine derivates along with the MCA moiety (donors of fluorescence) were introduced into a proteinase 3 (PR3) substrate with a C-terminal ANB-NH2 that serves as a fluorescence acceptor. Five substrates with general formula X-Tyr-Tyr-Abu-ANB-NH2 were synthesized, and their kinetic parameters against proteinase 3 were determined. The highest kcat/KM value, 1.5 × 106 M-1 s-1, was obtained for (Pyr)Box-Ala-Tyr-Tyr-Abu-NH 2 where (Pyr)Box-Ala stands for N-methylpyrrole benzoxazole-l- alanine. Titration of this peptide with proteinase 3 resulted in measurable fluorescence at an enzyme amount equal to 29 pmol. This substrate was selected used to detect quantifiable levels of proteinase 3 in serum samples, including those of normal subjects. For all c-ANCA-positive samples (diagnosed Wegener granulomatosis), a significant increase of PR3 concentration was observed. Wegener granulomatosis is a severe autoimmune disease causing inflammation of the blood vessels. Our results clearly show that this substrate can be used for the construction of a very reliable, inexpensive, and easy to use diagnostic test for PR3 determination.
