PXD063880-1

PXD063880 is an original dataset announced via ProteomeXchange.

Title	Investigating protein dynamics with millisecond precision using droplet microfluidic hydrogen/deuterium 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. Millisecond droplet HDX processors were used to measure forward exchange and demonstrate that D2O labelling is the rate-limiting step, in essence defining 10 milliseconds as the minimum practical incubation time. 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. Calcium binding increases D2O accessibility to the linker region, reporting the flexibility which enables the scissor-like motion of calmodulin for capturing proteins. The millisecond precision of droplet microfluidic HDX paves the way to advance our understanding of protein structural dynamics.
HostingRepository	PRIDE
AnnounceDate	2025-06-23
AnnouncementXML	Submission_2025-06-23_07:28:14.392.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: 9913;
ModificationList	No PTMs are included in the dataset
Instrument	Xevo G2-XS QTof

Revision	Datetime	Status	ChangeLog Entry
0	2025-05-13 01:09:03	ID requested
⏵ 1	2025-06-23 07:28:14	announced

Dataset with its publication pending

ProteomeXchange project tag: Hydrogen Deuterium Exchange (HDX-MS)

submitter keyword: calmodulin,HDX-MS, droplet microfluidics

Dr Eamonn Reading
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

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2025/06/PXD063880

PRIDE project URI

• PRIDE
  1. PXD063880
     1. Label: PRIDE project
     2. Name: Investigating protein dynamics with millisecond precision using droplet microfluidic hydrogen/deuterium exchange