Girard, Eric and Lopes, Pedro and Spoerner, Michael and Dhaussy, Anne-Claire and Prange, Thierry and Kalbitzer, Hans Robert and Colloc'h, Nathalie (2024) High Pressure Promotes Binding of the Allosteric Inhibitor Zn²⁺-Cyclen in Crystals of Activated H-Ras. CHEMISTRY-A EUROPEAN JOURNAL, 30 (35). e202400304. ISSN 0947-6539, 1521-3765
Full text not available from this repository. (Request a copy)Abstract
In this work, we experimentally investigate the potency of high pressure to drive a protein toward an excited state where an inhibitor targeted for this state can bind. Ras proteins are small GTPases cycling between active GTP-bound and inactive GDP-bound states. Various states of GTP-bound Ras in active conformation coexist in solution, amongst them, state 2 which binds to effectors, and state 1, weakly populated at ambient conditions, which has a low affinity for effectors. Zn2+-cyclen is an allosteric inhibitor of Ras protein, designed to bind specifically to the state 1. In H-Ras(wt).Mg2+.GppNHp crystals soaked with Zn2+-cyclen, no binding could be observed, as expected in the state 2 conformation which is the dominant state at ambient pressure. Interestingly, Zn2+-cyclen binding is observed at 500 MPa pressure, close to the nucleotide, in Ras protein that is driven by pressure to a state 1 conformer. The unknown binding mode of Zn2+-cyclen to H-Ras can thus be fully characterized in atomic details. As a more general conjunction from our study, high pressure x-ray crystallography turns out to be a powerful method to induce transitions allowing drug binding in proteins that are in low-populated conformations at ambient conditions, enabling the design of specific inhibitors.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | MACROMOLECULAR CRYSTALLOGRAPHY; PROTEIN CRYSTALLOGRAPHY; CONFORMATIONAL DYNAMICS; EFFECTOR INTERACTION; STRUCTURAL DYNAMICS; GTP HYDROLYSIS; EXCITED-STATE; K-RAS; NMR; MECHANISM; allosterism; drug design; high pressure crystallography HPMX; Ras protein; in crystallo transition |
| Subjects: | 500 Science > 540 Chemistry & allied sciences 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie > Prof. Dr. Dr. Hans Robert Kalbitzer |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 14 Jan 2026 05:59 |
| Last Modified: | 14 Jan 2026 05:59 |
| URI: | https://pred.uni-regensburg.de/id/eprint/64945 |
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