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  1. HRaDeX: high-resolution HDXโ€“MS analysis made simple โš™๏ธ๐Ÿงฌ
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../../

  • ๐Ÿ”— Try it yourself (no excuses ๐Ÿ˜‰)
  • ๐ŸŽง Audio summary
  • ๐Ÿ”ฌ What is this about?
  • ๐Ÿšจ The core limitation
  • ๐Ÿง  The idea behind HRaDeX
    • ๐Ÿงฉ Overlapping peptides
    • โš™๏ธ Kinetic modeling
  • ๐ŸŽจ Visualizing protein dynamics
  • โš™๏ธ How the workflow works
  • โš™๏ธ Two resolution strategies
    • 1๏ธโƒฃ Shortest peptide
    • 2๏ธโƒฃ Weighted average
  • ๐Ÿ“Š Performance & validation
  • ๐Ÿงฌ What you can actually do with it
  • ๐Ÿš€ Why this matters
  • โš ๏ธ Limitations
  • ๐Ÿ’š BioGenies perspective

HRaDeX: high-resolution HDXโ€“MS analysis made simple โš™๏ธ๐Ÿงฌ

HaDeX
publications
A new R package and web server (HRaDeX) enabling high-resolution analysis of HDXโ€“MS data by reconstructing residue-level deuterium uptake rates from peptide-level measurements.
Author

BioGenies Lab

Published

May 19, 2025

Keywords

HDX-MS, protein dynamics, structural proteomics, HRaDeX, mass spectrometry, protein folding, bioinformatics tools


๐Ÿ“Œ Project highlights

  • โš™๏ธ New tool: HRaDeX (R package + web server)
  • ๐Ÿงฌ Enables high-resolution HDXโ€“MS analysis
  • ๐Ÿ” Reconstructs residue-level dynamics from peptide data
  • ๐Ÿ“Š Uses extended kinetic modeling (ZS model)
  • ๐ŸŽฏ Validated on 4000+ peptides across multiple datasets

๐ŸŽ‰ New paper out! This one is all about turning messy HDXโ€“MS data into something actually interpretable ๐Ÿ˜„

๐Ÿ‘‰ HRaDeX: R Package and Web Server for Computing High-Resolution Deuterium Uptake Rates for HDXโ€“MS Data


๐Ÿ”— Try it yourself (no excuses ๐Ÿ˜‰)

HRaDeX is available as both a web application and an R package, allowing flexible use across workflows, from quick exploration to large-scale analysis.

  • ๐ŸŒ Web server
  • ๐Ÿ”ฌ Comparative server
  • ๐Ÿ’ป GitHub (R package)

๐ŸŽง Audio summary

HDXโ€“MS + peptide overlap + kinetic modeling = ๐Ÿ˜ตโ€๐Ÿ’ซ

๐Ÿ‘‰ So yes, this one also deserves a quick audio breakdown ๐ŸŽง

Your browser does not support the audio element.


๐Ÿ”ฌ What is this about?

HRaDeX is a bioinformatics tool for analyzing hydrogenโ€“deuterium exchange mass spectrometry (HDXโ€“MS) data, enabling reconstruction of protein dynamics at near residue-level resolution.

HDXโ€“MS measures how proteins exchange hydrogen with deuterium over time, revealing:

  • structural stability
  • folding dynamics
  • solvent accessibility

๐Ÿ‘‰ But standard analysis is limited to peptide-level resolution, averaging signals across multiple residues.


๐Ÿšจ The core limitation

Classic HDXโ€“MS:

  • โŒ loses residue-level detail
  • โŒ averages overlapping signals
  • โŒ makes precise interpretation difficult

๐Ÿ‘‰ This is where HRaDeX comes in.


๐Ÿง  The idea behind HRaDeX

HRaDeX reconstructs high-resolution information using:

๐Ÿงฉ Overlapping peptides

  • combines multiple fragments covering the same region

โš™๏ธ Kinetic modeling

  • fits uptake curves using extended ZS model
  • separates:
    • fast
    • intermediate
    • slow exchange components

๐Ÿ‘‰ Each peptide becomes a mixture of dynamic populations.


๐ŸŽจ Visualizing protein dynamics

One of the nicest features:

๐Ÿ‘‰ RGB color encoding of exchange rates

  • ๐Ÿ”ด fast (flexible)
  • ๐ŸŸข intermediate
  • ๐Ÿ”ต slow (structured)

โžก๏ธ giving a single-view map of dynamics across the sequence


โš™๏ธ How the workflow works

According to the workflow:

  1. โœ… Filter invalid data
  2. ๐ŸงŠ Detect โ€œno exchangeโ€ regions
  3. ๐Ÿ“ˆ Fit kinetic models
  4. ๐ŸŽฏ Select best model (BIC)
  5. ๐Ÿ”— Aggregate overlapping peptides
  6. ๐ŸŽจ Map results to sequence/structure

โš™๏ธ Two resolution strategies

HRaDeX provides:

1๏ธโƒฃ Shortest peptide

  • assigns values from smallest fragment

2๏ธโƒฃ Weighted average

  • integrates all overlapping peptides

๐Ÿ‘‰ The combination enables higher resolution than standard HDXโ€“MS


๐Ÿ“Š Performance & validation

Tested extensively:

  • โœ”๏ธ >4000 peptides
  • ๐Ÿ“‰ very low fitting error (RSS)
  • ๐ŸŽฏ ~4โ€“7% RMSE in reconstructed uptake

๐Ÿ‘‰ Meaning: HRaDeX reliably reproduces experimental HDX behavior


๐Ÿงฌ What you can actually do with it

  • map flexible vs stable regions
  • detect binding interfaces
  • study allosteric effects
  • compare protein states

๐Ÿ‘‰ Example: binding-induced conformational changes clearly visible in case studies


๐Ÿš€ Why this matters

This work bridges a major gap:

๐Ÿ‘‰ from peptide-level data โ†’ residue-level interpretation

It enables:

  • better structural insight
  • more precise protein dynamics analysis
  • improved reproducibility

โš ๏ธ Limitations

  • depends on peptide coverage
  • requires good experimental design
  • resolution limited if overlap is low

๐Ÿ‘‰ Still a major step forward compared to standard pipelines


๐Ÿ’š BioGenies perspective

This work fits directly into our focus on:

  • protein dynamics ๐Ÿงฌ
  • structural interpretation โš™๏ธ
  • computational biology ๐Ÿ”

๐Ÿ‘‰ and reinforces a key idea better models = more information from the same data

 

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