Novel internally quenched substrate of the trypsin-like subunit of 20S eukaryotic proteasome

Natalia Gruba, Magdalena Wysocka, Magdalena Brzezińska, Dawid Debowski, Krzysztof Rolka, Nathaniel I Martin, Adam Lesner*

*Corresponding author for this work

    Research output: Contribution to journalArticleAcademicpeer-review

    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.

    Original languageEnglish
    Pages (from-to)38-45
    Number of pages8
    JournalAnalytical Biochemistry
    Volume508
    DOIs
    Publication statusPublished - 1 Sept 2016

    Keywords

    • 20S Eukaryotic proteasome
    • Internally quenched peptides
    • Fluorescence

    Fingerprint

    Dive into the research topics of 'Novel internally quenched substrate of the trypsin-like subunit of 20S eukaryotic proteasome'. Together they form a unique fingerprint.

    Cite this