Spin‐Labeled Riboswitch Synthesized from a Protected TPA Phosphoramidite Building Block

The nitroxide TPA (2,2,5,5‐tetramethyl‐pyrrolin‐1‐oxyl‐3‐acetylene) is an excellent spin label for EPR studies of RNA. Previous synthetic methods, however, are complicated and require special equipment. Herein, we describe a uridine derived phosphoramidite with a photocaged TPA unit attached. The li...

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Detalles Bibliográficos
Autores principales: Kaiser, Frank, Endeward, Burkhard, Collauto, Alberto, Scheffer, Ute, Prisner, Thomas F., Göbel, Michael W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2022
Materias:
Research Articles
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9804336/
https://www.ncbi.nlm.nih.gov/pubmed/35903916
http://dx.doi.org/10.1002/chem.202201822
Descripción
Sumario:The nitroxide TPA (2,2,5,5‐tetramethyl‐pyrrolin‐1‐oxyl‐3‐acetylene) is an excellent spin label for EPR studies of RNA. Previous synthetic methods, however, are complicated and require special equipment. Herein, we describe a uridine derived phosphoramidite with a photocaged TPA unit attached. The light sensitive 2‐nitrobenzyloxymethyl group can be removed in high yield by short irradiation at 365 nm. Based on this approach, a doubly spin‐labeled 27mer neomycin sensing riboswitch was synthesized and studied by PELDOR. The overall thermal stability of the fold is not much reduced by TPA. In‐line probing nevertheless detected changes in local mobility.

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