Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
7557493 | Analytical Biochemistry | 2016 | 8 Pages |
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/KMÂ =Â 934,000Â Mâ1Â sâ1) and (ii) ABZ-Val-Val-Ser-GNF-Ala-Met-Gly-Tyr(3-NO2)-NH2 (kcat/KMÂ =Â 1,980,000Â Mâ1Â sâ1). Both compounds were efficiently hydrolyzed by the 20S proteasome at picomolar concentrations, demonstrating significant selectivity over other proteasome entities.
Keywords
ACCN,N,N′,N′-tetramethyl-O-(benzotriazol-1-yl)uronium tetrafluoroborateDIPCIANBTBTUPR3ABZHOBtAmCRP-HPLCTFA4-dimethylaminopyridineDMAPDMFSDS1-hydroxybenzotriazole2-aminobenzoic acid3-nitro-l-tyrosineN,N′-diisopropylcarbodiimideN,N-diisopropylethylamineTrifluoroacetic acidFluorescent Resonance Energy TransferFRETdimethylformamidesodium dodecyl sulfateubiquitin–proteasome systemFluorescenceDIPEAproteinase 3reverse phase high-performance liquid chromatographyUPS
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Natalia Gruba, Magdalena Wysocka, Magdalena BrzeziÅska, Dawid Debowski, Krzysztof Rolka, Nathaniel I. Martin, Adam Lesner,