Cross‐Polarization Schemes for Improved Heteronuclear Transfers Involving Labile Protons in Biomolecular Solution NMR

Abstract

AbstractINEPT‐based experiments are widely used for 1H→15N transfers, but often fail when involving labile protons due to solvent exchanges. J‐based cross polarization (CP) strategies offer a more efficient alternative to perform such transfers, particularly when leveraging the Hwater HN exchange process to boost the 1H→15N transfer process. This leveraging, however, demands the simultaneous spin‐locking of both Hwater and HN protons by a strong 1H RF field, while fulfilling the γHB1,H=γNB1,N Hartmann‐Hahn matching condition. Given the low value of γN/γH, however, these demands are often incompatible—particularly when experiments are executed by the power‐limited cryogenic probes used in contemporary high field NMR. The present manuscript discusses CP alternatives that can alleviate this limitation, and evaluates their performance on urea, amino acids, and intrinsically disordered proteins. These alternatives include new CP variants based on frequency‐swept and phase‐modulated pulses, designed to simultaneously fulfill the aforementioned conflicting conditions. Their performances vis‐à‐vis current options are theoretically analyzed with Liouville‐space simulations, and experimentally tested with double and triple resonance transfer experiments.