Hyperpolarizing DNA Nucleobases via NMR Signal Amplification by Reversible Exchange

The present work investigates the potential for enhancing the NMR signals of DNA nucleobases by parahydrogen-based hyperpolarization. Signal amplification by reversible exchange (SABRE) and SABRE in Shield Enables Alignment Transfer to Heteronuclei (SABRE-SHEATH) of selected DNA nucleobases is demonstrated with the enhancement (epsilon) of H-1, N-15, and/or C-13 spins in 3-methyladenine, cytosine, and 6-O-guanine. Solutions of the standard SABRE homogenous catalyst Ir(1,5-cyclooctadeine)(1,3-bis(2,4,6-trimethylphenyl)imidazolium)Cl (IrIMes) and a given nucleobase in deuterated ethanol/water solutions yielded low H-1 epsilon values (<= 10), likely reflecting weak catalyst binding. However, we achieved natural-abundance enhancement of N-15 signals for 3-methyladenine of similar to 3300 and similar to 1900 for the imidazole ring nitrogen atoms. H-1 and N-15 3-methyladenine studies revealed that methylation of adenine affords preferential binding of the imidazole ring over the pyrimidine ring. Interestingly, signal enhancements (epsilon similar to 240) of both N-15 atoms for doubly labelled cytosine reveal the preferential binding of specific tautomer(s), thus giving insight into the matching of polarization-transfer and tautomerization time scales. C-13 enhancements of up to nearly 50-fold were also obtained for this cytosine isotopomer. These efforts may enable the future investigation of processes underlying cellular function and/or dysfunction, including how DNA nucleobase tautomerization influences mismatching in base-pairing.

Automated summary

This study examines the potential for enhancing NMR signals of DNA nucleobases using parahydrogen-based hyperpolarization. Signal amplification by reversible exchange (SABRE) and SABRE in Shield Enables Alignment Transfer to Heteronuclei (SABRE-SHEATH) are demonstrated for selected DNA nucleobases. Low H-1 epsilon values (<10) were obtained for solutions of the standard SABRE catalyst and a given nucleobase, likely indicating weak catalyst binding. However, significant natural-abundance enhancement of N-15 signals was achieved for 3-methyladenine and cytosine. These findings provide insight into nucleobase tautomerization and may facilitate the investigation of cellular function and dysfunction.