An expanded allosteric network in PTP1B by multitemperature crystallography, fragment screening, and covalent tethering

DOI: 10.7554/eLife.36307

Authors: Daniel A. Keedy (University of California, San Francisco) , Zachary B. Hill (University of California, San Francisco) , Justin T. Biel (University of California, San Francisco) , Emily Kang (University of California, San Francisco) , T. Justin Rettenmaier (University of California, San Francisco) , Jose Brandao-Neto (Diamond Light Source) , Nicholas M. Pearce (Utrecht University) , Frank Von Delft (Diamond Light Source) , James A Wells (University of California, San Francisco) , James Fraser (University of California, San Francisco)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Elife , VOL 7

State: Published (Approved)
Published: June 2018
Diamond Proposal Number(s): 15751

Abstract: Allostery is an inherent feature of proteins, but it remains challenging to reveal the mechanisms by which allosteric signals propagate. A clearer understanding of this intrinsic circuitry would afford new opportunities to modulate protein function. Here we have identified allosteric sites in protein tyrosine phosphatase 1B (PTP1B) by combining multiple-temperature X-ray crystallography experiments and structure determination from hundreds of individual small-molecule fragment soaks. New modeling approaches reveal 'hidden' low-occupancy conformational states for protein and ligands. Our results converge on allosteric sites that are conformationally coupled to the active-site WPD loop and are hotspots for fragment binding. Targeting one of these sites with covalently tethered molecules or mutations allosterically inhibits enzyme activity. Overall, this work demonstrates how the ensemble nature of macromolecular structure, revealed here by multitemperature crystallography, can elucidate allosteric mechanisms and open new doors for long-range control of protein function.

Journal Keywords: allostery; X-ray crystallography; conformational heterogeneity; ligand binding; phosphatase

Diamond Keywords: Diabetes

Subject Areas: Biology and Bio-materials, Medicine


Instruments: I04-1-Macromolecular Crystallography (fixed wavelength)

Other Facilities: Advanced Light Source

Added On: 12/06/2018 11:13

Discipline Tags:

Non-Communicable Diseases Health & Wellbeing Structural biology Drug Discovery Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)