Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
7558883 | Analytical Biochemistry | 2014 | 24 Pages |
Abstract
Internally quenched cathepsin L (Cat L) substrate ABZ-Bip-Arg-Ala-Gln-Tyr(3-NO2)-NH2 with high specificity constant (kcat/KMÂ =Â 2.6Â ÃÂ 107Â Mâ1Â sâ1) was synthesized. The resultant compound displayed high selectivity over other members of the cathepsin family (B, S, X, V, C, K, H, F, D, and A). Activity of Cat L at picomolar (pM) concentrations was found using this substrate. Moreover, it was established that the presence of the selective Cat L inhibitor suppressed the proteolysis of the substrate to a non-detectable level. Incubation of the synthesized compound with a cell lysate of healthy and cancer cell lines indicated significant differences in Cat L activity. Based on the obtained results, it is proposed that this substrate could be used for selective monitoring of Cat L activity in biological systems.
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Monika ÅÄgowska, Magdalena Wysocka, Timo Burster, MichaÅ PikuÅa, Krzysztof Rolka, Adam Lesner,