An improved double-tuned and isotope-filtered pulse scheme based on a pulsed field gradient and a wide-band inversion shaped pulse.

We have developed an improved isotope-filtered pulse scheme in combination with a double-tuned filter, a hyperbolic secant inversion pulse, and a z-filter with a pulsed field gradient. These filtering pulse schemes have been incorporated into several one-, two-, and three-dimensional experiments, which were applied to the (13)C/(15)N uniformly labeled N-terminal SH3 domain of Grb2 complexed with the unlabeled Sos-derived peptide. The proton resonances of the Sos-derived peptide were unambiguously assigned using isotope-filtered DQF-COSY, TOCSY and NOESY spectra. Furthermore, in the isotope-filtered, isotope-edited 3D NOESY spectrum, intermolecular NOEs between the labeled protein and the unlabeled peptide could be identified. Through these applications, we demonstrate the high filtering efficiency of the presented pulse scheme.