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Journal article
An NMR‐Based Biosensor to Measure Stereospecific Methionine Sulfoxide Reductase Activities in Vitro and in Vivo
Chemistry : a European journal, Vol.26(65), pp.14838-14843
20/Nov/2020
PMID: 32501570
Abstract
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. Simultaneous activity measurements of MSRA and MSRB enzymes by NMR. CarMetOx is a synthetic biosensor to monitor the reduction of oxidized methionine residues by methionine sulfoxide reductase (MSR) enzymes. Based on NMR detection of R‐ and S‐ diastereoisomers of the isotope‐labelled reporter, stereospecific activities of class A (R‐) and class B (S‐) MSRs can be measured simultaneously in vitro and in vivo.
Details
- Title
- An NMR‐Based Biosensor to Measure Stereospecific Methionine Sulfoxide Reductase Activities in Vitro and in Vivo
- Creators
- Carolina Sánchez‐López - Instituto de Biología Molecular y Celular de RosarioNatalia Labadie - Instituto de Biología Molecular y Celular de RosarioVerónica A Lombardo - National University of RosarioFranco A Biglione - Instituto de Biología Molecular y Celular de RosarioBruno Manta - Brigham and Women's HospitalReeba Susan Jacob - 972WIS_INST___120Vadim N Gladyshev - Harvard Medical SchoolSalim Abdelilah‐Seyfried - University of PotsdamPhilipp Selenko - 972WIS_INST___120Andres Binolfi (Corresponding Author) - Instituto de Biología Molecular y Celular de Rosario
- Resource Type
- Journal article
- Publication Details
- Chemistry : a European journal, Vol.26(65), pp.14838-14843; 20/Nov/2020
- Number of pages
- 6
- Language
- English
- DOI
- https://doi.org/10.1002/chem.202002645
- PMID
- 32501570
- Grant note
- 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.
- Record Identifier
- 993248020303596
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