Cyclic Ion Mobility for Hydrogen/Deuterium Exchange-Mass Spectrometry Applications

Area of Science:

• Biophysical Chemistry
• Analytical Chemistry
• Structural Biology

Background:

• Hydrogen/deuterium exchange-mass spectrometry (HDX-MS) is crucial for studying protein dynamics at peptide resolution.
• Data quality in HDX-MS relies heavily on peptide identification and monitoring.
• Integral membrane proteins (IMPs) present challenges in HDX-MS due to low sequence coverage and data redundancy.

Purpose of the Study:

• To evaluate the performance of cyclic ion mobility (cIM) coupled with HDX-MS for analyzing integral membrane proteins (IMPs).
• To develop a novel data processing strategy for multipass cIM data.
• To assess the impact of cIM on peptide identification and data quality in HDX-MS workflows.

Main Methods:

• Utilized a SELECT SERIES Cyclic IM spectrometer for HDX-MS analysis of four detergent-solubilized IMP samples.
• Employed both one-pass and multipass cIM routines.
• Developed and applied a novel processing strategy for multipass cIM data.

Main Results:

• Coupling cIM with HDX-MS demonstrated enhanced performance for IMP analysis.
• One-pass and multipass cIM routines identified unique peptide populations.
• Combined peptide output increased by 31% to 222% compared to previous instrumentation (SYNAPT G2-Si).

Conclusions:

• The integration of cIM into HDX-MS workflows significantly improves peptide coverage and data quality for IMPs.
• The developed processing strategy effectively handles multipass cIM data.
• This novel HDX-MS workflow offers potential for more accurate and faster analysis of complex biological systems.