TY - JOUR
T1 - An NMR-Based Biosensor to Measure Stereospecific Methionine Sulfoxide Reductase Activities in Vitro and in Vivo
AU - Sánchez-López, Carolina
AU - Labadie, Natalia
AU - Lombardo, Verónica A.
AU - Biglione, Franco A.
AU - Manta, Bruno
AU - Gladyshev, Vadim N.
AU - Abdelilah-Seyfried, Salim
AU - Selenko, Philipp
AU - Binolfi, Andres
N1 - A.B. and V.A.L. acknowledge CONICET, ANPCyT (PICT 2017‐1241) and Fundación IBR for financial support. P.S. is a recipient of an ERC Consolidator Grant #647474 (NeuroInCellNMR). F.A.B. acknowledges CONICET for doctoral fellowship. C.S.L. acknowledges SECTEI CDMX, Mexico for a post‐doctoral fellowship. We thank Dr. Luciano Abriata and Dr. François‐Xavier Theillet for helpful discussions, Sebastian Graziati for assistance with fish handling and Alejandro Gago and Andrea Coscia for maintenance of the NMR facility.
PY - 2020/11/20
Y1 - 2020/11/20
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85092443331&partnerID=8YFLogxK
U2 - 10.1002/chem.202002645
DO - 10.1002/chem.202002645
M3 - مقالة
SN - 0947-6539
VL - 26
SP - 14838
EP - 14843
JO - Chemistry - A European Journal
JF - Chemistry - A European Journal
IS - 65
ER -