Abstract
Low-concentration photochemically induced dynamic polarization (LC-photo-CIDNP) enables the spectroscopic analysis of biomolecules containing the amino acids Trp and Tyr at sub-micromolar concentration in solution. Typical LC-photo-CIDNP pulse sequences involving 1H-13C correlation, however, perform well in the case of aromatic resonances but display a relatively poor signal-to-noise ratio for 13Cα and 13Cβ resonances. Here, we develop a novel pulse sequence denoted as 13C perturbation-recovered selective-pulse photo-CINDP enhanced reverse INEPT, or 13C PRESPRINT, tailored to the LC-photo-CIDNP analysis of 1H-13Cα pairs. Our method, which is based on full suppression of 1-bond Cα-C' scalar-coupling evolution during the constant-time delay, results into a sensitivity improvement by a factor of 2. The enhanced performance of this pulse sequence enabled us to improve the analysis of LC-photo-CIDNP laser-power dependence at very low (200 nM) sample concentration. An improved theoretical model, developed to quantitatively describe this laser-power dependence, shows excellent agreement with our 13C PRESPRINT experimental data.
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