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- W4285226021 abstract "Hydrogen deuterium exchange coupled to mass spectrometry (HDX-MS) is a valuable technique to investigate the dynamics of protein systems. The approach compares the deuterium uptake of protein backbone amides under multiple conditions to characterize protein conformation and interaction. HDX-MS is versatile and can be applied to diverse ligands, however, challenges remain when it comes to exploring complexes containing nucleic acids. In this chapter, we present procedures for the optimization and application of HDX-MS to studying RNA-binding proteins and use the RNA helicase Mtr4 as a demonstrative example. We highlight considerations in designing on-exchange, bottom-up, comparative studies on proteins with RNA. Our protocol details preliminary testing and optimization of experimental parameters. Difficulties arising from the inclusion of RNA, such as signal repression and sample carryover, are addressed. We discuss how chromatography parameters can be adjusted depending on the issues presented by the RNA, emphasizing reproducible peptide recovery in the absence and presence of RNA. Methods for visualization of HDX data integrated with statistical analysis are also reviewed with examples. These protocols can be applied to future studies of various RNA-protein complexes." @default.
- W4285226021 created "2022-07-14" @default.
- W4285226021 creator A5024626437 @default.
- W4285226021 creator A5038528104 @default.
- W4285226021 creator A5088977916 @default.
- W4285226021 creator A5091097798 @default.
- W4285226021 date "2022-01-01" @default.
- W4285226021 modified "2023-10-17" @default.
- W4285226021 title "Using hydrogen-deuterium exchange mass spectrometry to characterize Mtr4 interactions with RNA" @default.
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- W4285226021 doi "https://doi.org/10.1016/bs.mie.2022.04.002" @default.
- W4285226021 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/35965017" @default.
- W4285226021 hasPublicationYear "2022" @default.