5V8O

Discovery of a high affinity inhibitor of cGAS

5V8O の概要

• エントリーDOI: 10.2210/pdb5v8o/pdb
• 関連するPDBエントリー: 5V8H 5V8J 5V8N
• 分子名称: Cyclic GMP-AMP synthase, ZINC ION, 5-phenyltetrazolo[1,5-a]pyrimidin-7-ol, ... (4 entities in total)
• 機能のキーワード: cgas, sting, cgamp, transferase-transferase inhibitor complex, transferase/transferase inhibitor
• 由来する生物種: Homo sapiens (Human)
• 細胞内の位置: Cytoplasm, cytosol . Note=(Microbial infection) Upon infection with virulent M: Q8N884
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 85580.22

構造登録者

Hall, J. (登録日: 2017-03-22, 公開日: 2017-09-27, 最終更新日: 2024-03-06)

主引用文献

Hall, J.,Brault, A.,Vincent, F.,Weng, S.,Wang, H.,Dumlao, D.,Aulabaugh, A.,Aivazian, D.,Castro, D.,Chen, M.,Culp, J.,Dower, K.,Gardner, J.,Hawrylik, S.,Golenbock, D.,Hepworth, D.,Horn, M.,Jones, L.,Jones, P.,Latz, E.,Li, J.,Lin, L.L.,Lin, W.,Lin, D.,Lovering, F.,Niljanskul, N.,Nistler, R.,Pierce, B.,Plotnikova, O.,Schmitt, D.,Shanker, S.,Smith, J.,Snyder, W.,Subashi, T.,Trujillo, J.,Tyminski, E.,Wang, G.,Wong, J.,Lefker, B.,Dakin, L.,Leach, K.
Discovery of PF-06928215 as a high affinity inhibitor of cGAS enabled by a novel fluorescence polarization assay.
PLoS ONE, 12:e0184843-e0184843, 2017

PubMed Abstract: Cyclic GMP-AMP synthase (cGAS) initiates the innate immune system in response to cytosolic dsDNA. After binding and activation from dsDNA, cGAS uses ATP and GTP to synthesize 2', 3' -cGAMP (cGAMP), a cyclic dinucleotide second messenger with mixed 2'-5' and 3'-5' phosphodiester bonds. Inappropriate stimulation of cGAS has been implicated in autoimmune disease such as systemic lupus erythematosus, thus inhibition of cGAS may be of therapeutic benefit in some diseases; however, the size and polarity of the cGAS active site makes it a challenging target for the development of conventional substrate-competitive inhibitors. We report here the development of a high affinity (KD = 200 nM) inhibitor from a low affinity fragment hit with supporting biochemical and structural data showing these molecules bind to the cGAS active site. We also report a new high throughput cGAS fluorescence polarization (FP)-based assay to enable the rapid identification and optimization of cGAS inhibitors. This FP assay uses Cy5-labelled cGAMP in combination with a novel high affinity monoclonal antibody that specifically recognizes cGAMP with no cross reactivity to cAMP, cGMP, ATP, or GTP. Given its role in the innate immune response, cGAS is a promising therapeutic target for autoinflammatory disease. Our results demonstrate its druggability, provide a high affinity tool compound, and establish a high throughput assay for the identification of next generation cGAS inhibitors.

PubMed: 28934246
DOI: 10.1371/journal.pone.0184843

実験手法

X-RAY DIFFRACTION (3.1 Å)