dc.contributor.author | Cooley, R | |
dc.contributor.author | Kara, N | |
dc.contributor.author | Hui, NS | |
dc.contributor.author | Tart, J | |
dc.contributor.author | Roustan, C | |
dc.contributor.author | George, R | |
dc.contributor.author | Hancock, DC | |
dc.contributor.author | Binkowski, BF | |
dc.contributor.author | Wood, KV | |
dc.contributor.author | Ismail, M | |
dc.contributor.author | Downward, J | |
dc.date.accessioned | 2020-08-27T13:13:05Z | |
dc.date.issued | 2020-01-01 | |
dc.identifier.citation | Wellcome open research, 2020, 5 pp. 20 - ? | |
dc.identifier.issn | 2398-502X | |
dc.identifier.uri | https://repository.icr.ac.uk/handle/internal/4044 | |
dc.identifier.eissn | 2398-502X | |
dc.identifier.doi | 10.12688/wellcomeopenres.15675.1 | |
dc.description.abstract | Targeting the interaction of proteins with weak binding affinities or low solubility represents a particular challenge for drug screening. The NanoLuc â ® Binary Technology (NanoBiT â ®) was originally developed to detect protein-protein interactions in live mammalian cells. Here we report the successful translation of the NanoBit cellular assay into a biochemical, cell-free format using mammalian cell lysates. We show that the assay is suitable for the detection of both strong and weak protein interactions such as those involving the binding of RAS oncoproteins to either RAF or phosphoinositide 3-kinase (PI3K) effectors respectively, and that it is also effective for the study of poorly soluble protein domains such as the RAS binding domain of PI3K. Furthermore, the RAS interaction assay is sensitive and responds to both strong and weak RAS inhibitors. Our data show that the assay is robust, reproducible, cost-effective, and can be adapted for small and large-scale screening approaches. The NanoBit Biochemical Assay offers an attractive tool for drug screening against challenging protein-protein interaction targets, including the interaction of RAS with PI3K. | |
dc.format | Electronic-eCollection | |
dc.format.extent | 20 - ? | |
dc.language | eng | |
dc.language.iso | eng | |
dc.publisher | F1000 Research Ltd | |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0 | |
dc.title | Development of a cell-free split-luciferase biochemical assay as a tool for screening for inhibitors of challenging protein-protein interaction targets. | |
dc.type | Journal Article | |
dcterms.dateAccepted | 2020-01-29 | |
rioxxterms.versionofrecord | 10.12688/wellcomeopenres.15675.1 | |
rioxxterms.licenseref.uri | https://creativecommons.org/licenses/by/4.0 | |
rioxxterms.licenseref.startdate | 2020-01 | |
rioxxterms.type | Journal Article/Review | |
dc.relation.isPartOf | Wellcome open research | |
pubs.notes | Not known | |
pubs.organisational-group | /ICR | |
pubs.organisational-group | /ICR/Primary Group | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Cancer Biology | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Cancer Biology/Lung Cancer Group | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Cancer Therapeutics | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Cancer Therapeutics/Signal Transduction & Molecular Pharmacology | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Molecular Pathology | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Molecular Pathology/Lung Cancer Group | |
pubs.organisational-group | /ICR/Students | |
pubs.organisational-group | /ICR/Students/PhD and MPhil | |
pubs.organisational-group | /ICR/Students/PhD and MPhil/20/21 Starting Cohort | |
pubs.organisational-group | /ICR | |
pubs.organisational-group | /ICR/Primary Group | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Cancer Biology | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Cancer Biology/Lung Cancer Group | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Cancer Therapeutics | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Cancer Therapeutics/Signal Transduction & Molecular Pharmacology | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Molecular Pathology | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Molecular Pathology/Lung Cancer Group | |
pubs.organisational-group | /ICR/Students | |
pubs.organisational-group | /ICR/Students/PhD and MPhil | |
pubs.organisational-group | /ICR/Students/PhD and MPhil/20/21 Starting Cohort | |
pubs.publication-status | Published | |
pubs.volume | 5 | |
pubs.embargo.terms | Not known | |
icr.researchteam | Signal Transduction & Molecular Pharmacology | |
icr.researchteam | Lung Cancer Group | |
dc.contributor.icrauthor | Cooley, Rachel | |