Native dynamics and allosteric responses in PTP1B probed by high-resolution HDX-MS

Area of Science:

• Biophysics
• Structural Biology
• Pharmacology

Background:

• Protein tyrosine phosphatase 1B (PTP1B) is a key therapeutic target for metabolic diseases and cancer.
• Understanding PTP1B's conformational dynamics and allostery is crucial for developing effective allosteric inhibitors.

Approach:

• Employed high-resolution hydrogen-deuterium exchange mass spectrometry (HDX-MS) to map PTP1B's solution dynamics.
• Generated HDX-MS maps for apo-PTP1B and in the presence of active-site and allosteric inhibitors.

Key Points:

• HDX-MS revealed flexible loops and distinct dynamic regions in apo-PTP1B, adding value to crystal structure-based predictions.
• Inhibitor binding induced significant, long-range alterations in PTP1B's dynamics, distinct for active-site vs. allosteric inhibitors.
• Observed allosteric effects extended over 35 Å from ligand binding sites, highlighting PTP1B's complex conformational landscape.

Conclusions:

• HDX-MS is a powerful technique for elucidating protein dynamics and allosteric mechanisms.
• Findings provide critical insights into PTP1B allostery, supporting the rational design of novel therapeutics.
• Integration of HDX-MS with other methods can accelerate the development of PTP1B-targeted drugs.