Počet záznamů: 1
Enzymatic Synthesis of 3′-5′, 3′-5′ Cyclic Dinucleotides, Their Binding Properties to the Stimulator of Interferon Genes Adaptor Protein, and Structure/Activity Correlations
- 1.0549413 - ÚOCHB 2022 RIV US eng J - Článek v odborném periodiku
Novotná, Barbora - Holá, Lucie - Staś, Monika - Gutten, Ondrej - Smola, Miroslav - Zavřel, Martin - Vavřina, Zdeněk - Buděšínský, Miloš - Liboska, Radek - Chevrier, Florian - Dobiaš, Juraj - Bouřa, Evžen - Rulíšek, Lubomír - Birkuš, Gabriel
Enzymatic Synthesis of 3′-5′, 3′-5′ Cyclic Dinucleotides, Their Binding Properties to the Stimulator of Interferon Genes Adaptor Protein, and Structure/Activity Correlations.
Biochemistry. Roč. 60, č. 48 (2021), s. 3714-3727. ISSN 0006-2960. E-ISSN 1520-4995
Grant CEP: GA MŠMT(CZ) EF16_019/0000729; GA ČR(CZ) GA20-08772S
Institucionální podpora: RVO:61388963
Klíčová slova: cytosolic DNA sensor * c-di-GMP * structural analysis
Obor OECD: Biochemistry and molecular biology
Impakt faktor: 3.321, rok: 2021
Způsob publikování: Omezený přístup
https://doi.org/10.1021/acs.biochem.1c00692
The 3′–5′, 3′–5′ cyclic dinucleotides (3′3′CDNs) are bacterial second messengers that can also bind to the stimulator of interferon genes (STING) adaptor protein in vertebrates and activate the host innate immunity. Here, we profiled the substrate specificity of four bacterial dinucleotide synthases from Vibrio cholerae (DncV), Bacillus thuringiensis (btDisA), Escherichia coli (dgcZ), and Thermotoga maritima (tDGC) using a library of 33 nucleoside-5′-triphosphate analogues and then employed these enzymes to synthesize 24 3′3′CDNs. The STING affinity of CDNs was evaluated in cell-based and biochemical assays, and their ability to induce cytokines was determined by employing human peripheral blood mononuclear cells. Interestingly, the prepared heterodimeric 3′3′CDNs bound to the STING much better than their homodimeric counterparts and showed similar or better potency than bacterial 3′3′CDNs. We also rationalized the experimental findings by in-depth STING-CDN structure–activity correlations by dissecting computed interaction free energies into a set of well-defined and intuitive terms. To this aim, we employed state-of-the-art methods of computational chemistry, such as quantum mechanics/molecular mechanics (QM/MM) calculations, and complemented the computed results with the {STING:3′3′c-di-ara-AMP} X-ray crystallographic structure. QM/MM identified three outliers (mostly homodimers) for which we have no clear explanation of their impaired binding with respect to their heterodimeric counterparts, whereas the R2 = 0.7 correlation between the computed ΔG′int_rel and experimental ΔTm’s for the remaining ligands has been very encouraging.
Trvalý link: http://hdl.handle.net/11104/0325428
Počet záznamů: 1