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

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*

*Corresponding author for this work

Research output: Contribution to JournalArticleAcademicpeer-review

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.

Original languageEnglish
Article numbere36307
JournaleLife
Volume7
DOIs
Publication statusPublished - 7 Jun 2018
Externally publishedYes

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    Keedy, D. A., Hill, Z. B., Biel, J. T., Kang, E., Rettenmaier, T. J., Brandão-Neto, J., ... Fraser, J. S. (2018). An expanded allosteric network in PTP1B by multitemperature crystallography, fragment screening, and covalent tethering. eLife, 7, [e36307]. https://doi.org/10.7554/eLife.36307