PXD063880 is an
original dataset announced via ProteomeXchange.
Dataset Summary
| Title | Droplet microfluidic hydrogen/deuterium exchange for investigating protein dynamics with millisecond precision exchange |
| Description | Hydrogen/deuterium exchange (HDX) methods for studying protein dynamics would benefit from millisecond-scale incubations to probe intrinsically disordered proteins, highly dynamic regions and conformation changes. Here we investigate droplet microfluidics for rapid mixing to trigger D2O labelling, uniform incubations and rapid droplet merging for acid quenching in advance of mass spectrometry. A surfactant-free merging approach combining expansion elements for synchronised droplet collision proved robust. The high diffusive flux of D2O and protons enable microsecond mixing to trigger and arrest D2O labelling, respectively, affording the possibility of single millisecond incubations. Droplet HDX processors were used to measure the fast uptake characteristics of a model peptide. Forward exchange measurements demonstrate D2O labelling to be the rate-limiting step, in essence defining 10 milliseconds as the minimum practical incubation time for proteins in typical physiological conditions. With the ability to access millisecond time scales the fast dynamics of calmodulin, a model of calcium-triggered allostery with rapid conformational switching, was investigated. At 10 milliseconds, we could observe significant deuterium uptake within the well-defined EF-hand motifs (Ca2+ binding sites). These findings demonstrate millisecond HDX enabled by droplet microfluidics allows areas of heightened plasticity to be detected within a stably folded protein.
Associated dataset: PXD077479 |
| HostingRepository | PRIDE |
| AnnounceDate | 2026-05-27 |
| AnnouncementXML | Submission_2026-05-27_08:38:51.899.xml |
| DigitalObjectIdentifier | |
| ReviewLevel | Peer-reviewed dataset |
| DatasetOrigin | Original dataset |
| RepositorySupport | Unsupported dataset by repository |
| PrimarySubmitter | Alistair Bailey |
| SpeciesList | scientific name: Bos taurus (Bovine); NCBI TaxID: NEWT:9913; |
| ModificationList | No PTMs are included in the dataset |
| Instrument | Xevo G2-XS QTof |
Dataset History
| Revision | Datetime | Status | ChangeLog Entry |
|---|
| 0 | 2025-05-13 01:09:03 | ID requested | |
| 1 | 2025-06-23 07:28:14 | announced | |
| 2 | 2026-04-22 01:23:30 | announced | 2026-04-22: Updated project metadata. |
| 3 | 2026-04-22 09:12:31 | announced | 2026-04-22: Updated project metadata. |
| ⏵ 4 | 2026-05-27 08:38:52 | announced | 2026-05-27: Updated project metadata. |
Publication List
| Dataset with its publication pending |
Keyword List
| ProteomeXchange project tag: Hydrogen Deuterium Exchange (HDX-MS) |
| submitter keyword: calmodulin,HDX-MS, droplet microfluidics |
Contact List
| Dr Eamonn |
|---|
| contact affiliation | School of Biological Sciences, Building 85, University of Southampton, Southampton, SO17 1BJ |
| contact email | E.Reading@soton.ac.uk |
| lab head | |
| Alistair Bailey |
|---|
| contact affiliation | University of Southampton |
| contact email | ab604@soton.ac.uk |
| dataset submitter | |
Full Dataset Link List
Dataset FTP location
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| PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD063880
- Label: PRIDE project
- Name: Droplet microfluidic hydrogen/deuterium exchange for investigating protein dynamics with millisecond precision exchange
