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.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1002/chem.202002645 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Small Molecule Screening Strategies from Lead Identification to Validation.
J Vis Exp
January 2023
Department of Cancer Biology, Chemical Biology Program, Dana-Farber Cancer Institute;
Festa, F., Labaer, J. Kinase inhibitor screening in self-assembled human protein microarrays.
View Article and Find Full Text PDF-Cryptophanes: macrocyclic compounds with optimized characteristics for the design of Xe NMR-based biosensors.
Phys Chem Chem Phys
October 2022
NIMBE, CEA, CNRS, Université de Paris Saclay, CEA Saclay, 91191 Gif-sur-Yvette, France.
A new water-soluble xenon host system with great promise for the Xe NMR-based biosensing approach is presented: the -cryptophane-222-hexacarboxylate. It compares favorably with its already known diastereomer, on the one hand, and with cucurbit[6]uril, on the other hand, in particular in terms of xenon binding constant and xenon in-out exchange, a key parameter for the efficiency of the most sensitive HyperCEST method.
View Article and Find Full Text PDFEnhancing the detection sensitivity of nanobody against aflatoxin B through structure-guided modification.
Int J Biol Macromol
January 2022
State Key Laboratory of Magnetic Resonance and Atomic Molecular Physics, Key Laboratory of Magnetic Resonance in Biological Systems, National Center for Magnetic Resonance in Wuhan, Wuhan Institute of Physics and Mathematics, Innovation Academy for Precision Measurement Science and Technology, Chinese Academy of Sciences - Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Hubei Optics Valley Laboratory, Wuhan 430071, China; University of Chinese Academy of Sciences, Beijing 100049, China.
Nanobodies (Nbs) have shown great potential in immunodetection of small-molecule contaminants in food and environmental monitoring. However, the limited knowledge of the mechanism of Nbs binding to small molecules has hampered the development of high-affinity Nbs and assay improvement. We previously reported two homologous nanobodies Nb26 and Nb28 specific to aflatoxin B (AFB), with the former exhibiting higher sensitivity in ELISA.
View Article and Find Full Text PDFPathological aggregation of the protein tau into insoluble aggregates is a hallmark of neurodegenerative diseases. The emergence of disease-specific tau aggregate structures termed tau strains, however, remains elusive. Here we show that full-length tau protein can be aggregated in the absence of co-factors into seeding-competent amyloid fibrils that sequester RNA.
View Article and Find Full Text PDFAn NMR-Based Biosensor to Measure Stereospecific Methionine Sulfoxide Reductase Activities in Vitro and in Vivo*.
Chemistry
November 2020
Instituto de Biología Molecular y Celular de Rosario (IBR-CONICET-UNR), Ocampo y Esmeralda, 2000, Rosario, Argentina.
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.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!