Dipolar recoupling in solid-state NMR is an essential method for establishing correlations between nuclei that are close in space.Inapplications on protein samples,the traditional experiments like ramped and adiabatic DCP suffer from the fact that dipolar recoupling occurs only within alimited volume of the sample.This selection is dictated by the radiofrequency (rf) field inhomogeneity profile of the excitation solenoidal coil. We employo ptimal control strategies to design dipolar recoupling sequences with substantially larger responsive volume and increased sensitivity.W eshow that it is essential to compensate for additional temporal modulations induced by sample rotation in as patially inhomogeneous rf field. Suchm odulations interfere with the pulse sequence and decrease its performance.U sing large-scale optimizations we developed pulse schemes for magnetization transfer from amide nitrogen to carbonyl (NCO) as well as aliphatic carbons (NCA). Our experiments yield as ignal intensity increased by af actor of 1.5 and 2.0 for NCA and NCO transfers, respectively,c ompared to conventional ramped DCP sequences.C onsistent results were obtained using several biological samples and NMR instruments.
Experimentalsensitivityisthemajorobstacleinapplicationsof solid state nuclear magnetic resonance (NMR) in structural biology. [1] In studies of protein samples,c orrelations of chemical shifts between amide nitrogen and carbon atoms of the protein backbone are essential for resonance assignments. [2] Fort his purpose,d ipolar recoupling methods are employed. In the basic variant of the double cross polarization (DCP) experiment, [3] ar esonant radiofrequency( rf) field is applied to both the 15 Nand 13 Cnuclei which interferes with the spatial rotation of the sample (magic angle spinning, MAS) and re-introduces the direct dipole-dipole interaction. Foranefficient magnetization transfer, the rf amplitudes have to satisfy the Hartmann-Hahn (HH) condition [4] (w N AE w C ¼ nw R ,w here w R =2p is the sample spinning frequency, w N and w C are the rf amplitudes on the 15 Na nd 13 C channels,r espectively,e xpressed in rad s À1 ,a nd n ¼ AE1; AE2). However,d ue to rf field inhomogeneities imposed by the NMR coil the HH condition cannot be fulfilled for the whole volume of the sample and it leads to aloss of sensitivity.T he most widely used experiments today are the ramped and adiabatic DCP [5] in which the rf amplitude on one of the rf channels is modulated and swept through the HH condition. Experimentally,t he efficiency of the DCP experiment is evaluated as the ratio of the signal intensities of the 13 CC P-MAS spectrum and the DCP spectrum. Thecurrent standard efficiency of the ramped/adiabatic DCP experiment is 30-40 %when arotor is completely filled with sample,and about 65 %w hen the sample is restricted to the central slice of the rotor where the rf field is expected to be homogeneous. [6] It is obvious that the experiment does not work equally well for the whole sample volume and it is estimated that 30-50 %of the...