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dc.contributor.authorLi, Chan
dc.contributor.authorReddy, Tummala Rama Krishna
dc.contributor.authorFischer, Peter M.
dc.contributor.authorDekker, Lodewijk V.
dc.date.accessioned2013-01-11T15:29:05Z
dc.date.available2013-01-11T15:29:05Z
dc.date.issued2010
dc.identifier.citationLi , C , Reddy , T R K , Fischer , P M & Dekker , L V 2010 , ' Cy5-labeled S100A10 tracer used to identify inhibitors of the protein interaction with annexin A2 ' , Assay and Drug Development Technologies , vol. 8 , no. 1 , pp. 85-95 . https://doi.org/10.1089/adt.2009.0218
dc.identifier.issn1557-8127
dc.identifier.otherPURE: 1374826
dc.identifier.otherPURE UUID: d9055105-7ecf-4a14-9fe8-53d09353f0d4
dc.identifier.otherScopus: 77649098499
dc.identifier.urihttp://hdl.handle.net/2299/9595
dc.description.abstractProtein–protein interactions are increasingly of interest as targets in small-molecule drug discovery. The interaction between the Ca2+- and phospholipid-binding protein Annexin A2 and its binding partner S100A10 has been implicated in angiogenesis and cancer metastasis. Here, we present a methodology to screen for inhibitors of this protein interaction. We developed a Cy5-labeled S100A10 tracer and showed by circular dichroism spectroscopy that the secondary structure is indistinguishable from that of non-labeled S100A10. This tracer was used to develop a binding assay based upon fluorescence resonance energy transfer to a Cy3-labeled Annexin A2 peptide ligand. The binding parameters matched those for unlabeled components as observed by equilibrium dialysis, which we determined separately, as well as those determined by isothermal titration calorimetry. Binding of labeled and unlabeled peptide was specific and mutually competitive. We used this assay for screening a small compound library derived by computational interrogation of the S100A10-binding pocket. Hits were obtained with IC50 values in range of the IC50 of the cognate Annexin A2 peptide ligand. Hits were subjected to an exact parallel assay measuring an unrelated protein–protein interaction (antigen–antibody). In this way, we identified genuine hits that inhibited the interaction between S100A10 and Annexin A2 but do not affect the fluorescence readout. These compounds are potentially of interest as candidates for further analysis and medical chemistry exploration. The simple assay format described here can be employed in early-stage exploration of other protein–protein interaction targetsen
dc.language.isoeng
dc.relation.ispartofAssay and Drug Development Technologies
dc.titleCy5-labeled S100A10 tracer used to identify inhibitors of the protein interaction with annexin A2en
dc.contributor.institutionDepartment of Pharmacy
dc.contributor.institutionSchool of Life and Medical Sciences
dc.contributor.institutionHealth & Human Sciences Research Institute
dc.contributor.institutionMedicinal and Analytical Chemistry
dc.description.statusPeer reviewed
rioxxterms.versionVoR
rioxxterms.versionofrecordhttps://doi.org/10.1089/adt.2009.0218
rioxxterms.typeJournal Article/Review
herts.preservation.rarelyaccessedtrue


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