Combined Protein- and Ligand-Observed NMR Workflow to Screen Fragment Cocktails against Multiple Proteins: A Case Study Using Bromodomains.
Combined Protein- and Ligand-Observed NMR Workflow to Screen Fragment Cocktails against Multiple Proteins: A Case Study Using Bromodomains.
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DOI：
10.3390/molecules25173949
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发表时间：
2020-08-29
期刊：
影响因子：
--
通讯作者：
Pomerantz WCK
中科院分区：
文献类型：
--
作者：
Johnson JA;Olson NM;Tooker MJ;Bur SK;Pomerantz WCK
As fragment-based drug discovery has become mainstream, there has been an increase in various screening methodologies. Protein-observed 19F (PrOF) NMR and 1H CPMG NMR are two fragment screening assays that have complementary advantages. Here, we sought to combine these two NMR-based assays into a new screening workflow. This combination of protein- and ligand-observed experiments allows for a time- and resource-efficient multiplexed screen of mixtures of fragments and proteins. PrOF NMR is first used to screen mixtures against two proteins. Hit mixtures for each protein are identified then deconvoluted using 1H CPMG NMR. We demonstrate the benefit of this fragment screening method by conducting the first reported fragment screens against the bromodomains of BPTF and Plasmodium falciparum (Pf) GCN5 using 467 3D-enriched fragments. The hit rates were 6%, 5% and 4% for fragments binding BPTF, PfGCN5, and fragments binding both proteins, respectively. Select hits were characterized, revealing a broad range of affinities from low µM to mM dissociation constants. Follow-up experiments supported a low-affinity second binding site on PfGCN5. This approach can be used to bias fragment screens towards more selective hits at the onset of inhibitor development in a resource- and time-efficient manner.
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BPTF保持染色质的可及性和乳腺干细胞的自我更新能力。
影响因子：
5.9
作者：
Frey WD;Chaudhry A;Slepicka PF;Ouellette AM;Kirberger SE;Pomerantz WCK;Hannon GJ;Dos Santos CO
通讯作者：
Dos Santos CO
DOI：
10.1002/chem.201704169
发表时间：
2017-10-26
期刊：
Chemistry (Weinheim an der Bergstrasse, Germany)
影响因子：
--
作者：
Foley DJ;Craven PGE;Collins PM;Doveston RG;Aimon A;Talon R;Churcher I;von Delft F;Marsden SP;Nelson A
通讯作者：
Nelson A
影响因子：
5.6
作者：
Chen, Dan;Ranganathan, Anirudh;Carlsson, Jens
通讯作者：
Carlsson, Jens
影响因子：
10.3
作者：
Dar, Altaf A.;Nosrati, Mehdi;Kashani-Sabet, Mohammed
通讯作者：
Kashani-Sabet, Mohammed
DOI：
10.1021/ci0200467
发表时间：
2003-01-01
期刊：
JOURNAL OF CHEMICAL INFORMATION AND COMPUTER SCIENCES
影响因子：
--
作者：
Feher, M;Schmidt, JM
通讯作者：
Schmidt, JM