An NMR-Based Biosensor to Measure Stereospecific Methionine Sulfoxide Reductase Activities in Vitro and in Vivo*.

Autor: Sánchez-López C; Instituto de Biología Molecular y Celular de Rosario (IBR-CONICET-UNR), Ocampo y Esmeralda, 2000, Rosario, Argentina., Labadie N; Instituto de Biología Molecular y Celular de Rosario (IBR-CONICET-UNR), Ocampo y Esmeralda, 2000, Rosario, Argentina., Lombardo VA; Instituto de Biología Molecular y Celular de Rosario (IBR-CONICET-UNR), Ocampo y Esmeralda, 2000, Rosario, Argentina.; Centro de Estudios Interdisciplinarios (CEI), Universidad Nacional de Rosario, 2000, Rosario, Argentina., Biglione FA; Instituto de Biología Molecular y Celular de Rosario (IBR-CONICET-UNR), Ocampo y Esmeralda, 2000, Rosario, Argentina., Manta B; Division of Genetics, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, 02115, USA.; Facultad de Medicina, Departamento de Bioquímica and Centro de Investigaciones Biomédicas, Universidad de la República, CP, 11800, Montevideo, Uruguay., Jacob RS; Department of Biological Regulation, Weizmann Institute of Science, 234 Herzl Street, 761000, Rehovot, Israel., Gladyshev VN; Division of Genetics, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, 02115, USA., Abdelilah-Seyfried S; Institute of Biochemistry and Biology, Potsdam University, 14476, Potsdam, Germany.; Institute of Molecular Biology, Hannover Medical School, 30625, Hannover, Germany., Selenko P; Department of Biological Regulation, Weizmann Institute of Science, 234 Herzl Street, 761000, Rehovot, Israel., Binolfi A; Instituto de Biología Molecular y Celular de Rosario (IBR-CONICET-UNR), Ocampo y Esmeralda, 2000, Rosario, Argentina.; Plataforma Argentina de Biología EstructuralyMetabolómica (PLABEM), Ocampo y Esmeralda, 2000, Rosario, Argentina.
Jazyk: angličtina
Zdroj: Chemistry (Weinheim an der Bergstrasse, Germany) [Chemistry] 2020 Nov 20; Vol. 26 (65), pp. 14838-14843. Date of Electronic Publication: 2020 Oct 14.
DOI: 10.1002/chem.202002645
Abstrakt: Oxidation of protein methionines to methionine-sulfoxides (MetOx) is associated with several age-related diseases. In healthy cells, MetOx is reduced to methionine by two families of conserved methionine sulfoxide reductase enzymes, MSRA and MSRB that specifically target the S- or R-diastereoisomers of methionine-sulfoxides, respectively. To directly interrogate MSRA and MSRB functions in cellular settings, we developed an NMR-based biosensor that we call CarMetOx to simultaneously measure both enzyme activities in single reaction setups. We demonstrate the suitability of our strategy to delineate MSR functions in complex biological environments, including cell lysates and live zebrafish embryos. Thereby, we establish differences in substrate specificities between prokaryotic and eukaryotic MSRs and introduce CarMetOx as a highly sensitive tool for studying therapeutic targets of oxidative stress-related human diseases and redox regulated signaling pathways.
(© 2020 Wiley-VCH GmbH.)
Databáze: MEDLINE
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