Synthesis of <sup>15</sup>N-Enriched Pseudouridine Derivatives

Desaulniers, Jean-Paul; Ksebati, Bashar; Chow, Christine S.

doi:10.1021/ol035496e

Cited by 6 publications

(2 citation statements)

References 22 publications

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“…[4] A previous synthetic route for the production of 3-15 N-labelled Ψ and its methylated derivative made use of a protected Cnucleoside precursor, in which the N3 position was nitrated and treated with 15 N-ammonia to give the 15 N3 isotopologue. [5] A drawback of this approach was the moderate 15 N labelling degree of ca. 45 %.…”

Section: Introductionmentioning

confidence: 99%

“…With the introduction of NMR active heteronuclei the effect of Ψ on the RNA's folding landscape can be thoroughly investigated by applying state of the art dynamic NMR experiments, such as relaxation dispersion (RD) and chemical exchange saturation transfer (CEST) pulse programs [4] . A previous synthetic route for the production of 3‐ 15 N‐labelled Ψ and its methylated derivative made use of a protected C‐nucleoside precursor, in which the N3 position was nitrated and treated with 15 N‐ammonia to give the 15 N3 isotopologue [5] . A drawback of this approach was the moderate 15 N labelling degree of ca.…”

Section: Introductionmentioning

confidence: 99%

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Efficient Synthetic Access to Stable Isotope Labelled Pseudouridine Phosphoramidites for RNA NMR Spectroscopy

Glänzer,

Pfeiffer,

Ribar

et al. 2024

Chemistry A European J

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Here we report the efficient synthetic access to 13C/15N‐labelled pseudouridine phosphoramidites, which were incorporated into a binary H/ACA box guide RNA/product complex comprising 77 nucleotides (nts) in total and into a 75 nt E. coli tRNAGly. The stable isotope (SI) labelled pseudouridines were produced via a highly efficient chemo‐enzymatic synthesis. 13C/15N labelled uracils were produced via chemical synthesis and enzymatically converted to 5′‐monophosphate pseudouridine (ΨMP) by using YeiN, a Ψ‐5′‐monophosphate C‐glycosidase. Removal of the 5′‐phosphate group yielded the desired pseudouridine nucleoside (Ψ), which was transformed into a phosphoramidite building suitable for RNA solid phase synthesis. A Ψ ‐building block carrying both a 13C and a 15N label was incorporated into a product RNA and the complex formation with a 63 nt H/ACA box RNA could be observed via NMR. Furthermore, the SI labelled pseudouridine building block was used to determine imino proton bulk water exchange rates of a 75 nt E. coli tRNAGly CCmnm5U, identifying the TΨC‐loop 5‐methyluridine as a modifier of the exchange rates. The efficient synthetic access to SI labelled Ψ building blocks will allow the solution and solid‐state NMR spectroscopic studies of Ψ containing RNAs and will facilitate the mass spectrometric analysis of Ψ‐modified nucleic acids.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%