Protein-Ligand Interactions in the STING Binding Site Probed by Rationally Designed Single-Point Mutations: Experiment and Theory

0554896 - BTÚ 2022 RIV US eng J - Journal Article
Vavrina, Z. - Gutten, O. - Smola, M. - Zavrel, M. - Tehrani, Zahra Aliakbar - Charvat, V. - Kožíšek, M. - Bouřa, E. - Birkus, G. - Rulíšek, L.
Protein-Ligand Interactions in the STING Binding Site Probed by Rationally Designed Single-Point Mutations: Experiment and Theory.
Biochemistry. Roč. 60, č. 8 (2021), s. 607-620. ISSN 0006-2960
Institutional support: RVO:86652036
Keywords : cyclic di-gmp * interferon genes * dna sensor * dinucleotide
OECD category: Biochemistry and molecular biology
Impact factor: 3.321, year: 2021
Method of publishing: Limited access
https://pubs.acs.org/doi/10.1021/acs.biochem.0c00949

STING protein (stimulator of interferon genes) plays an important role in the innate immune system. A number of potent compounds regulating its activity have been reported, mostly derivatives of cyclic dinucleotides (CDNs), natural STING agonists. Here, we aim to provide complementary information to large-scale ligand-profiling studies by probing the importance of STING-CDN protein-ligand interactions on the protein side. We examined in detail six typical CDNs each in complex with 13 rationally devised mutations in STING: S162A, S162T, Y167F, G230A, R232K, R232H, A233L, A233I, R238K, T263A, T263S, R293Q, and G230A/R293Q The mutations switch on and off various types of protein-ligand interactions: pi-pi stacking, hydrogen bonding, ionic pairing, and nonpolar contacts. We correlated experimental data obtained by differential scanning fluorimetry, X-ray crystallography, and isothermal titration calorimetry with theoretical calculations. This enabled us to provide a mechanistic interpretation of the differences in the binding of representative CDNs to STING. We observed that the G230A mutation increased the thermal stability of the protein-ligand complex, indicating an increased level of ligand binding, whereas R238K and Y167F led to a complete loss of stabilization (ligand binding). The effects of the other mutations depended on the type of ligand (CDN) and varied, to some extent. A very good correlation (R-2 = 0.6) between the experimental binding affinities and interaction energies computed by quantum chemical methods enabled us to explain the effect of the studied mutations in detail and evaluate specific interactions quantitatively. Our work may inspire development of high-affinity ligands against the common STING haplotypes by targeting the key (sometimes non-intuitive) protein-ligand interactions.
Permanent Link: http://hdl.handle.net/11104/0329535