A Basis for Reduced Chemical Library Inhibition of Firefly Luciferase Obtained from Directed Evolution

被引：59

作者：

Auld, Douglas S. ^{[1
]}

Zhang, Ya-Qin ^{[1
]}

Southall, Noel T. ^{[1
]}

Rai, Ganesha ^{[1
]}

Landsman, Marc ^{[1
]}

MacLure, Jennifer ^{[1
]}

Langevin, Daniel ^{[1
]}

Thomas, Craig J. ^{[1
]}

Austin, Christopher P. ^{[1
]}

Inglese, James ^{[1
]}

机构：

[1] NIH, Chem Genom Ctr, Bethesda, MD 20892 USA

来源：

JOURNAL OF MEDICINAL CHEMISTRY | 2009年 / 52卷 / 05期

关键词：

THROUGHPUT SCREENING ASSAYS; PROTEIN-KINASE; BIOLUMINESCENT ASSAYS; IDENTIFICATION; TRANSCREENER(TM); FLUORESCENCE; ACTIVATION; DISCOVERY; DRUGABILITY; MECHANISM;

D O I：

10.1021/jm8014525

中图分类号：

R914 [药物化学];

学科分类号：

100701 ;

摘要：

We measured the "druggability" of the ATP-dependent luciferase derived from the firefly Photuris pennsylvanica that was optimized using directed evolution (Ultra-Glo, Promega). Quantitative high-throughput screening (qHTS) was used to determine IC(50)S of 198899 samples against a formulation of Ultra-Glo luciferase (Kinase-Glo). We found that only 0.1 % of the Kinase-Glo inhibitors showed an IC(50) < 10 mu M compared to 0.9% found from a previous qHTS against the firefly luciferase from Photinus pyralis (lucPpy). Further, the maximum affinity identified in the lucPpy qHTS was 50 nM, while for Kinase-Glo this value increased to 600 nM. Compounds with interactions stretching outside the luciferin binding pocket were largely lost with Ultra-Glo luciferase. Therefore, Ultra-Glo luciferase will show less compound interference when used as an ATP sensor compared to lucPpy. This study demonstrates the power of large-scale quantitative analysis of structure-activity relationships (> 100K compounds) in addressing important questions such as a target's druggability.

引用

页码：1450 / 1458

页数：9

共 46 条

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