Identification of acylation products in SHAPE Chemistry

Chaoqi Lin (1) , Salomé Poyer (2) , Loussiné Zargarian (3) , Jean-Yves Salpin (2) , Philippe Fossé (3) , Olivier Mauffret (3) , Juan Xie (1)

1 PPSM - Laboratoire de Photophysique et Photochimie Supramoléculaires et Macromoléculaires
2 LAMBE - Laboratoire Analyse et Modélisation pour la Biologie et l'Environnement
3 LBPA - Laboratoire de Biologie et de Pharmacologie Appliquée

SHAPE chemistry (selective 2’-hydroxyl acylation analyzed by primer extension) has been developed to specifically target flexible nucleotides (often unpaired nucleotides) independently to their purine or pyrimidine nature for RNA secondary structure determination. However, to the best of our knowledge, the structure of 2’-O-acylation products has never been confirmed by NMR or X-ray data. We have realized the acylation reactions between cNMP and NMIA under SHAPE chemistry conditions and identified the acylation products using standard NMR spectroscopy and LC-MS/MS experiments. For cAMP and cGMP, the major acylation product is the 2’-O-acylated compound (> 99 %). A trace amount of N-acylated cAMP has also been identified by LC-UV-MS². While for cCMP, the isolated acylation products are composed of 96 % of 2’-O-acylated, 4 % of N,O-diacylated, and trace amount of N-acylated compounds. In addition, the characterization of the major 2’-O-acylated compound by NMR showed slight differences in the conformation of the acylated sugar between the three cyclic nucleotides. This interesting result should be useful to explain some unexpected reactivity of the SHAPE chemistry.

Mots clés: NMR; cNMP; Acylation reaction; LC-UV-MS²; SHAPE chemistry; RNA

Domaines: Chimie-Physique [physics.chem-ph]; Biochimie [q-bio.BM]

Chaoqi Lin, Salomé Poyer, Loussiné Zargarian, Jean-Yves Salpin, Philippe Fossé, et al.. Identification of acylation products in SHAPE Chemistry. Bioorganic and Medicinal Chemistry Letters, 2017, 27 (11), pp.2506-2509. ⟨10.1016/j.bmcl.2017.03.096⟩. ⟨hal-01637313⟩