Application of temperature-responsive HIS-tag fluorophores to differential scanning fluorimetry screening of small molecule libraries.
Application of temperature-responsive HIS-tag fluorophores to differential scanning fluorimetry screening of small molecule libraries.
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DOI：
10.3389/fphar.2022.1040039
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发表时间：
2022
影响因子：
5.6
通讯作者：
中科院分区：
文献类型：
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Differential scanning fluorimetry is a rapid and economical biophysical technique used to monitor perturbations to protein structure during a thermal gradient, most often by detecting protein unfolding events through an environment-sensitive fluorophore. By employing an NTA-complexed fluorophore that is sensitive to nearby structural changes in histidine-tagged protein, a robust and sensitive differential scanning fluorimetry (DSF) assay is established with the specificity of an affinity tag-based system. We developed, optimized, and miniaturized this HIS-tag DSF assay (HIS-DSF) into a 1536-well high-throughput biophysical platform using the Borrelial high temperature requirement A protease (BbHtrA) as a proof of concept for the workflow. A production run of the BbHtrA HIS-DSF assay showed a tight negative control group distribution of Tm values with an average coefficient of variation of 0.51% and median coefficient of variation of compound Tm of 0.26%. The HIS-DSF platform will provide an additional assay platform for future drug discovery campaigns with applications in buffer screening and optimization, target engagement screening, and other biophysical assay efforts.
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影响因子：
5.9
作者：
Cho H;Choi Y;Min K;Son JB;Park H;Lee HH;Kim S
通讯作者：
Kim S
影响因子：
3.6
作者：
Kariu T;Yang X;Marks CB;Zhang X;Pal U
通讯作者：
Pal U
DOI：
10.1007/978-1-0716-0954-5_16
发表时间：
2021-01-01
期刊：
PLANT CHEMICAL GENOMICS, 2 EDITION
影响因子：
--
作者：
Li, Xiaohui;Zhang, Chunhua
通讯作者：
Zhang, Chunhua
影响因子：
3.4
作者：
Zhuang X;Yang X;Altieri AS;Nelson DC;Pal U
通讯作者：
Pal U
DOI：
10.1007/978-1-0716-0163-1_4
发表时间：
2020-01-01
期刊：
TARGETING ENZYMES FOR PHARMACEUTICAL DEVELOPMENT: METHODS AND PROTOCOLS
影响因子：
--
作者：
Baljinnyam, Bolormaa;Ronzetti, Michael;Simeonov, Anton
通讯作者：
Simeonov, Anton