Development of a High-Throughput Fluorescence Polarization Assay to Identify Novel Ligands of Glutamate Carboxypeptidase II
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Alquicer, Glenda | - |
dc.contributor.author | Sedlak, David | - |
dc.contributor.author | Byun, Youngjoo | - |
dc.contributor.author | Pavlicek, Jiri | - |
dc.contributor.author | Stathis, Marigo | - |
dc.contributor.author | Rojas, Camilo | - |
dc.contributor.author | Slusher, Barbara | - |
dc.contributor.author | Pomper, Martin G. | - |
dc.contributor.author | Bartunek, Petr | - |
dc.contributor.author | Barinka, Cyril | - |
dc.date.accessioned | 2021-09-06T16:15:04Z | - |
dc.date.available | 2021-09-06T16:15:04Z | - |
dc.date.created | 2021-06-18 | - |
dc.date.issued | 2012-09 | - |
dc.identifier.issn | 1087-0571 | - |
dc.identifier.uri | https://scholar.korea.ac.kr/handle/2021.sw.korea/107635 | - |
dc.description.abstract | Glutamate carboxypeptidase II (GCPII) is an important target for therapeutic and diagnostic interventions aimed at prostate cancer and neurologic disorders. Here we describe the development and optimization of a high-throughput screening (HTS) assay based on fluorescence polarization (FP) that facilitates the identification of novel scaffolds inhibiting GCPII. First, we designed and synthesized a fluorescence probe based on a urea-based inhibitory scaffold covalently linked to a Bodipy TMR fluorophore (TMRGlu). Next, we established and optimized conditions suitable for HTS and evaluated the assay robustness by testing the influence of a variety of physicochemical parameters (e.g., pH, temperature, time) and additives. Using known GCPII inhibitors, the FP assay was shown to be comparable to benchmark assays established in the field. Finally, we evaluated the FP assay by HTS of a 20 000-compound library. The novel assay presented here is robust, highly reproducible (Z' = 0.82), inexpensive, and suitable for automation, thus providing an excellent platform for HTS of small-molecule libraries targeting GCPII. | - |
dc.language | English | - |
dc.language.iso | en | - |
dc.publisher | SAGE PUBLICATIONS INC | - |
dc.subject | UREA-BASED INHIBITORS | - |
dc.subject | MEMBRANE ANTIGEN | - |
dc.subject | PROSTATE-CANCER | - |
dc.subject | SUBSTRATE-SPECIFICITY | - |
dc.subject | BRAIN | - |
dc.subject | EXPRESSION | - |
dc.subject | NAALADASE | - |
dc.subject | DIAGNOSIS | - |
dc.subject | DESIGN | - |
dc.subject | NAAG | - |
dc.title | Development of a High-Throughput Fluorescence Polarization Assay to Identify Novel Ligands of Glutamate Carboxypeptidase II | - |
dc.type | Article | - |
dc.contributor.affiliatedAuthor | Byun, Youngjoo | - |
dc.identifier.doi | 10.1177/1087057112451924 | - |
dc.identifier.scopusid | 2-s2.0-84864997955 | - |
dc.identifier.wosid | 000307543000003 | - |
dc.identifier.bibliographicCitation | JOURNAL OF BIOMOLECULAR SCREENING, v.17, no.8, pp.1030 - 1040 | - |
dc.relation.isPartOf | JOURNAL OF BIOMOLECULAR SCREENING | - |
dc.citation.title | JOURNAL OF BIOMOLECULAR SCREENING | - |
dc.citation.volume | 17 | - |
dc.citation.number | 8 | - |
dc.citation.startPage | 1030 | - |
dc.citation.endPage | 1040 | - |
dc.type.rims | ART | - |
dc.type.docType | Article | - |
dc.description.journalClass | 1 | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
dc.relation.journalResearchArea | Biochemistry & Molecular Biology | - |
dc.relation.journalResearchArea | Biotechnology & Applied Microbiology | - |
dc.relation.journalResearchArea | Chemistry | - |
dc.relation.journalWebOfScienceCategory | Biochemical Research Methods | - |
dc.relation.journalWebOfScienceCategory | Biotechnology & Applied Microbiology | - |
dc.relation.journalWebOfScienceCategory | Chemistry, Analytical | - |
dc.subject.keywordPlus | UREA-BASED INHIBITORS | - |
dc.subject.keywordPlus | MEMBRANE ANTIGEN | - |
dc.subject.keywordPlus | PROSTATE-CANCER | - |
dc.subject.keywordPlus | SUBSTRATE-SPECIFICITY | - |
dc.subject.keywordPlus | BRAIN | - |
dc.subject.keywordPlus | EXPRESSION | - |
dc.subject.keywordPlus | NAALADASE | - |
dc.subject.keywordPlus | DIAGNOSIS | - |
dc.subject.keywordPlus | DESIGN | - |
dc.subject.keywordPlus | NAAG | - |
dc.subject.keywordAuthor | fluorescence polarization | - |
dc.subject.keywordAuthor | high-throughput screening | - |
dc.subject.keywordAuthor | glutamate carboxypeptidase II | - |
dc.subject.keywordAuthor | prostate-specific membrane antigen | - |
dc.subject.keywordAuthor | metallopeptidase | - |
Items in ScholarWorks are protected by copyright, with all rights reserved, unless otherwise indicated.
(02841) 서울특별시 성북구 안암로 14502-3290-1114
COPYRIGHT © 2021 Korea University. All Rights Reserved.
Certain data included herein are derived from the © Web of Science of Clarivate Analytics. All rights reserved.
You may not copy or re-distribute this material in whole or in part without the prior written consent of Clarivate Analytics.