Synthesis of a 13 C-methylene-labeled isoleucine precursor as a useful tool for studying protein side-chain interactions and dynamics.

Autor: Höfurthner T; Christian Doppler Laboratory for High-Content Structural Biology and Biotechnology, Department of Structural and Computational Biology, Max Perutz Laboratories, University of Vienna, Campus Vienna Biocenter 5, 1030, Vienna, Austria.; Vienna Doctoral School in Chemistry (DoSChem), University of Vienna, Währingerstraße 42, 1090, Vienna, Austria., Toscano G; Christian Doppler Laboratory for High-Content Structural Biology and Biotechnology, Institute of Organic Chemistry, University of Vienna, Währingerstraße 38, 1090, Vienna, Austria.; Vienna Doctoral School in Chemistry (DoSChem), University of Vienna, Währingerstraße 42, 1090, Vienna, Austria., Kontaxis G; Department of Structural and Computational Biology, Max Perutz Laboratories, University of Vienna, Campus Vienna Biocenter 5, 1030, Vienna, Austria., Beier A; Christian Doppler Laboratory for High-Content Structural Biology and Biotechnology, Department of Structural and Computational Biology, Max Perutz Laboratories, University of Vienna, Campus Vienna Biocenter 5, 1030, Vienna, Austria., Mayer M; Boehringer Ingelheim RCV GmbH & Co. KG, Dr. Boehringer Gasse 5-11, 1121, Vienna, Austria., Geist L; Boehringer Ingelheim RCV GmbH & Co. KG, Dr. Boehringer Gasse 5-11, 1121, Vienna, Austria., McConnell DB; Boehringer Ingelheim RCV GmbH & Co. KG, Dr. Boehringer Gasse 5-11, 1121, Vienna, Austria., Weinstabl H; Boehringer Ingelheim RCV GmbH & Co. KG, Dr. Boehringer Gasse 5-11, 1121, Vienna, Austria., Lichtenecker R; Christian Doppler Laboratory for High-Content Structural Biology and Biotechnology, Institute of Organic Chemistry, University of Vienna, Währingerstraße 38, 1090, Vienna, Austria. roman.lichtenecker@univie.ac.at., Konrat R; Christian Doppler Laboratory for High-Content Structural Biology and Biotechnology, Department of Structural and Computational Biology, Max Perutz Laboratories, University of Vienna, Campus Vienna Biocenter 5, 1030, Vienna, Austria. robert.konrat@univie.ac.at.; Department of Structural and Computational Biology, Max Perutz Laboratories, University of Vienna, Campus Vienna Biocenter 5, 1030, Vienna, Austria. robert.konrat@univie.ac.at.
Jazyk: angličtina
Zdroj: Journal of biomolecular NMR [J Biomol NMR] 2024 Mar; Vol. 78 (1), pp. 1-8. Date of Electronic Publication: 2023 Oct 11.
DOI: 10.1007/s10858-023-00427-2
Abstrakt: In this study, we present the synthesis and incorporation of a metabolic isoleucine precursor compound for selective methylene labeling. The utility of this novel α-ketoacid isotopologue is shown by incorporation into the protein Brd4-BD1, which regulates gene expression by binding to acetylated histones. High quality single quantum 13 C- 1  H-HSQC were obtained, as well as triple quantum HTQC spectra, which are superior in terms of significantly increased 13 C-T 2 times. Additionally, large chemical shift perturbations upon ligand binding were observed. Our study thus proves the great sensitivity of this precursor as a reporter for side-chain dynamic studies and for investigations of CH-π interactions in protein-ligand complexes.
(© 2023. The Author(s).)
Databáze: MEDLINE
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