Multiplexed Random Peptide Library and Phospho-Specific Antibodies Facilitate Human Polo-Like Kinase 1 Inhibitor Screen

Tanaka, Kenji; Koresawa, Mitsuhiko; Iida, Masato; Fukasawa, Kazuhiro; Stec, Erica; Cassaday, Jason; Chase, Peter; Rickert, Keith; Hodder, Peter; Takagi, Toshimitsu; Komatani, Hideya

doi:10.1089/adt.2009.0212

Cited by 2 publications

(2 citation statements)

References 42 publications

(57 reference statements)

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“…The most extensively used VS methods are docking [11], pharmacophore [12], quantitative structure activity relationship (QSAR) [2,3,13], similarity searching [5], and machine learning [14]. These methods are often divided into structure-based virtual screening (SBVS) and ligand-based virtual screening (LBVS) depending on what is already known about a target and its ligands Fig.…”

Section: Brief Overview Of Extensive Used Virtual Screening Methodsmentioning

confidence: 99%

Virtual Screening Methods as Tools for Drug Lead Discovery from Large Chemical Libraries

Zhu²,

Liu³

et al. 2012

CMC

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Virtual screening methods have been developed and explored as useful tools for searching drug lead compounds from chemical libraries, including large libraries that have become publically available. In this review, we discussed the new developments in exploring virtual screening methods for enhanced performance in searching large chemical libraries, their applications in screening libraries of ~ 1 million or more compounds in the last five years, the difficulties in their applications, and the strategies for further improving these methods.

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Section: Brief Overview Of Extensive Used Virtual Screening Methodsmentioning

confidence: 99%

Virtual Screening Methods as Tools for Drug Lead Discovery from Large Chemical Libraries

Zhu²,

Liu³

et al. 2012

CMC

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“…Non-radiometric based platforms, measuring 1) phosphorylated product such as ELISAs, 10 time-resolved fluorometry (DELFIA™), 11 time-resolved fluorescence resonance energy transfer (TR-FRET), 12,13 and fluorescence polarization (FP), 14 or 2) ATP consumption such as kinetic coupled assays, 15 and luminescence-based assays, 16 have been developed and successfully implemented in many screening laboratories. The former platform relies on specific anti-phosphopeptide antibodies and/or the correct peptide sequence for the substrate.…”

Section: Introductionmentioning

confidence: 99%

Comparison of Luminescence ADP Production Assay and Radiometric Scintillation Proximity Assay for Cdc7 Kinase

Takagi

Shum²,

Parisi³

et al. 2011

CCHTS

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Several assay technologies have been successfully adapted and used in HTS to screen for protein kinase inhibitors; however, emerging comparative analysis studies report very low hit overlap between the different technologies, which challenges the working assumption that hit identification is not dependent on the assay method of choice. To help address this issue, we performed two screens on the cancer target, Cdc7-Dbf4 heterodimeric protein kinase, using a direct assay detection method measuring [33P]-phosphate incorporation into the substrate and an indirect method measuring residual ADP production using luminescence. We conducted the two screens under similar conditions, where in one, we measured [33P]-phosphate incorporation using scintillation proximity assay (SPA), and in the other, we detected luminescence signal of the ATP-dependent luciferase after regenerating ATP from residual ADP (LUM). Surprisingly, little or no correlation were observed between the positives identified by the two methods; at a threshold of 30% inhibition, 25 positives were identified in the LUM screen whereas the SPA screen only identified two positives, Tannic acid and Gentian violet, with Tannic acid being common to both. We tested 20 out of the 25 positive compounds in secondary confirmatory study and confirmed 12 compounds including Tannic acid as Cdc7-Dbf4 kinase inhibitors. Gentian violet, which was only positive in the SPA screen, inhibited luminescence detection and categorized as a false positive. This report demonstrates the strong impact in detection format on the success of a screening campaign and the importance of carefully designed confirmatory assays to eliminate those compounds that target the detection part of the assay.

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Multiplexed Random Peptide Library and Phospho-Specific Antibodies Facilitate Human Polo-Like Kinase 1 Inhibitor Screen

Cited by 2 publications

References 42 publications

Virtual Screening Methods as Tools for Drug Lead Discovery from Large Chemical Libraries

Virtual Screening Methods as Tools for Drug Lead Discovery from Large Chemical Libraries

Comparison of Luminescence ADP Production Assay and Radiometric Scintillation Proximity Assay for Cdc7 Kinase

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