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[JOVE] Time-resolved ElectroSpray Ionization Hydrogen-deuterium Exchange Mass Spectrometry for Study


Abstract:

Intrinsically disordered proteins (IDPs) have long been a challenge to structural biologists due to their lack of stable secondary structure elements. Hydrogen-Deuterium Exchange (HDX) measured at rapid time scales is uniquely suited to detect structures and hydrogen bonding networks that are briefly populated, allowing for the characterization of transient conformers in native ensembles. Coupling of HDX to mass spectrometry offers several key advantages, including high sensitivity, low sample consumption and no restriction on protein size. This technique has advanced greatly in the last several decades, including the ability to monitor HDX labeling times on the millisecond time scale. In addition, by incorporating the HDX workflow onto a microfluidic platform housing an acidic protease microreactor, we are able to localize dynamic properties at the peptide level. In this study, Time-Resolved ElectroSpray Ionization Mass Spectrometry (TRESI-MS) coupled to HDX was used to provide a detailed picture of residual structure in the tau protein, as well as the conformational shifts induced upon hyperphosphorylation.

Cristina Lento1, Shaolong Zhu1, Kerene A. Brown1, Ruth Knox1, Peter Liuni1, Derek J. Wilson1,2,3

1Department of Chemistry, York University, 2The Centre for Research in Mass Spectrometry, York University, 3The Centre for Research on Biomolecular Interactions, York University

Link: https://www.jove.com/video/55464/time-resolved-electrospray-ionization-hydrogen-deuterium-exchange

#04292017 #massspectrometry #protein #labonachip #microreactor

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