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

• DOI: 10.7554/elife.36307
• Publication Date: 2018-06-06T12:00:13Z
• Authors: Daniel A Keedy; Zachary B Hill; Justin T Biel; Emily Kang; T Justin Rettenmaier; José Brandão-Neto; Nicholas M Pearce; Frank von Delft; James A Wells; James S Fraser
• Keywords: Allosteric enzyme

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