Ligand Screening and Discovery using Cocktail Soaking and Automated MicroED.

IF 3.6 4区 医学 Q2 CHEMISTRY, MEDICINAL
ChemMedChem Pub Date : 2025-03-26 DOI:10.1002/cmdc.202500156
Jieye Lin, Marc J Gallenito, Johan Hattne, Tamir Gonen
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引用次数: 0

Abstract

Cocktail soaking using single-crystal X-ray diffraction (SC-XRD) previously allowed high-throughput crystallographic screening of ligands against protein targets. However, protein microcrystals are not amenable to this approach if they are too small to yield strong diffraction patterns. In this study, we developed a workflow integrating cocktail soaking with automated microcrystal electron diffraction (MicroED) to allow rapid ligand screening, structure determination, and binding analysis directly from microcrystals. This can improve the successful hit rate, because binding is often more efficient when smaller crystals are soaked in the ligand. The approach was validated with known ligands of thermolysin and identified novel binding interactions for ligands of proteinase K. The structures of multiple protein-ligand complexes, including ligands with weak binding affinities, could be solved rapidly. Their estimated relative binding affinities are in good agreement with previous work and independent microscale thermophoresis (MST) measurements.

利用鸡尾酒浸泡和自动微电子显微镜筛选和发现配体。
以前,使用单晶x射线衍射(SC-XRD)进行鸡尾酒浸泡可以对蛋白质靶点进行高通量晶体学筛选。然而,如果蛋白质微晶体太小而不能产生强烈的衍射图案,则不适合这种方法。在这项研究中,我们开发了一种将鸡尾酒浸泡与自动微晶体电子衍射(MicroED)相结合的工作流程,可以直接从微晶体中快速筛选配体,确定结构和结合分析。这可以提高成功命中率,因为当较小的晶体浸泡在配体中时,结合通常更有效。该方法与已知的热溶酶配体进行了验证,并发现了蛋白酶k配体的新的结合相互作用。多种蛋白质-配体复合物的结构,包括具有弱结合亲和力的配体,可以快速求解。他们估计的相对结合亲和力与先前的工作和独立的微尺度热泳术(MST)测量结果很好地一致。
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来源期刊
ChemMedChem
ChemMedChem 医学-药学
CiteScore
6.70
自引率
2.90%
发文量
280
审稿时长
1 months
期刊介绍: Quality research. Outstanding publications. With an impact factor of 3.124 (2019), ChemMedChem is a top journal for research at the interface of chemistry, biology and medicine. It is published on behalf of Chemistry Europe, an association of 16 European chemical societies. ChemMedChem publishes primary as well as critical secondary and tertiary information from authors across and for the world. Its mission is to integrate the wide and flourishing field of medicinal and pharmaceutical sciences, ranging from drug design and discovery to drug development and delivery, from molecular modeling to combinatorial chemistry, from target validation to lead generation and ADMET studies. ChemMedChem typically covers topics on small molecules, therapeutic macromolecules, peptides, peptidomimetics, and aptamers, protein-drug conjugates, nucleic acid therapies, and beginning 2017, nanomedicine, particularly 1) targeted nanodelivery, 2) theranostic nanoparticles, and 3) nanodrugs. Contents ChemMedChem publishes an attractive mixture of: Full Papers and Communications Reviews and Minireviews Patent Reviews Highlights and Concepts Book and Multimedia Reviews.
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