Simultaneous acquisition of [13C,15N]- and [15N,15N]-separated 4D gradient-enhanced NOESY spectra in proteins.

The simultaneous acquisition of a 4D gradient-enhanced and sensitivity-enhanced [13C,15N]/[15N,15N]-separated NOESY is presented for the 74-residue [13C,15N]-labeled N-terminal SH3 domain of mGrb2 complexed with a peptide fragment from mSOS-2 in 90% H2O. The method readily accommodates different 13C and 15N spectral widths, but requires that the same number of increments be collected for both 13C and 15N in the simultaneous dimension (F2). For purposes of display and analysis, the two 4D spectra can be deconvolved during the processing stage by the appropriate linear combination of separately stored FIDs. Compared to collecting each of these two 4D data sets separately, the presented method is a factor (2)1/2 more efficient in sensitivity per unit acquisition time. The interleaved nature of this method may also lead to improved peak registration between the two 4D spectra.