Journal of Physical Chemistry B, Vol.113, No.24, 8310-8318, 2009
H-1 Photo-CIDNP Enhancements in Heteronuclear Correlation NMR Spectroscopy
Photochemically induced dynamic nuclear polarization (photo-CIDNP) is usually employed as a probe of solvent exposure in biomolecular NMR. The potential of the photo-CIDNP effect for sensitivity enhancement, however, remains poorly explored. Here, we introduce H-1-photo-CIDNP in heteronuclear correlation spectroscopy at low laser irradiation power (1 W), and compare the sensitivity of various H-1-photo-CIDNP-enhanced-(HPE) H-1-N-15 heteronuclear correlation pulse sequences, including HSQC, HMQC, and SOFAST-HMQC, in terms of their ability to detect the Trp indole H-epsilon 1 resonance. Both Trp and the Trp-containing protein apoHmpH were analyzed using flavin mononucleotide as photosensitizer in aqueous solutions either containing or lacking urea. We find that H-1-N-15 photo-CIDNP-SOFAST-HMQC, denoted here as HPESOFAST-HMQC, yields a 2-fold higher signal-to-noise per unit time than the parent SOFAST-HMQC, for the solvent-exposed Trp of urea-unfolded apoHmpH. Thus, HPE-SOFAST-HMQC is the most sensitive heteronuclear correlation pulse sequence-for the detection of solvent-exposed Trp.