Novel internally quenched substrate of the trypsin-like subunit of 20S eukaryotic proteasome
Publication date
2016-09-01
Authors
Gruba, Natalia
Wysocka, Magdalena
Brzezińska, Magdalena
Debowski, Dawid
Rolka, Krzysztof
Martin, Nathaniel I
Lesner, Adam
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Supervisors
Document Type
Article
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Abstract
This article describes the synthesis, using combinatorial chemistry, of internally quenched substrates of the trypsin-like subunit of human 20S proteasome. Such substrates were optimized in both the nonprime and prime regions of the peptide chain. Two were selected as the most susceptible for proteasomal proteolysis with excellent kinetic parameters: (i) ABZ-Val-Val-Ser-Arg-Ser-Leu-Gly-Tyr(3-NO2)-NH2 (kcat/KM = 934,000 M(-1) s(-1)) and (ii) ABZ-Val-Val-Ser-GNF-Ala-Met-Gly-Tyr(3-NO2)-NH2 (kcat/KM = 1,980,000 M(-1) s(-1)). Both compounds were efficiently hydrolyzed by the 20S proteasome at picomolar concentrations, demonstrating significant selectivity over other proteasome entities.
Keywords
20S Eukaryotic proteasome, Internally quenched peptides, Fluorescence, Taverne
Citation
Gruba, N, Wysocka, M, Brzezińska, M, Debowski, D, Rolka, K, Martin, N I & Lesner, A 2016, 'Novel internally quenched substrate of the trypsin-like subunit of 20S eukaryotic proteasome', Analytical Biochemistry, vol. 508, pp. 38-45. https://doi.org/10.1016/j.ab.2015.08.019