Toward the discovery of new antifungal agents: The design and validation of a novel 2′P-RNA probe and high throughput screening assay against 2′-phosphotransferase Tpt1p

• Published in: Nucleic Acids Symposium Series Vol. 52; no. 1; pp. 475 - 476
• Main Authors: Lackey, Jeremy G., Ron, David, Damha, Masad J., Harding, Heather P.
• Format: Journal Article
• Language: English
• Published: Oxford University Press 01.09.2008
• ISSN: 0261-3166; 1746-8272
• DOI: 10.1093/nass/nrn241

Abstract We report the solid-phase synthesis of novel 2′P-RNA probes for use in fluorescence polarization (FP) ligand binding assays that screens for inhibitors of the yeast 2′- phosphotransferase Tpt1p. The probe was synthesized by utilizing silyl phosphoramidite chemistry and a phosphoramidite synthon containing an orthogonal (DMT) protecting group at its 2′-position. Regioselective removal of the 2′-DMT group and phosphitylation of the unmasked 2′-hydroxyl group afforded the desired 2′P-RNA sequence