4.4 Sequence-specific resonance assignments
NMR spectra were acquired at 20 °C on an 800 MHz NMR spectrometer equipped with a triple-resonance cryogenic probe. The following spectra (with measurement times) were acquired for sequence-specific resonance assignment of backbone and side-chain nuclei: 2D HSQC (0.6 h), 3D HNCO (4.2 h), 3D HNCACB (13 h), 3D CBCACONH (13 h), 3D HBHACONH (6.8 h), 3D (H)C(CO)NH TOCSY (10.4 h), 3D 13C-TOCSY-Methyl COSY (11.2 h), Reduced Dimensionality (RD) 3D HN(CA)NH (12.4 h), GFT (3,2)D HNHA (6 h). Data were processed with NMRPipe and analyzed using XEASY. Resonance assignments were obtained using a semi-automated approach in the program AUTOASSIGN. Sequence-specific resonance assignments were aided by amino acid selective unlabeling of Arg, Asn, Thr, Ser, Gly, and Ala and reduced dimensionality. The complete chemical shifts assignments have been deposited in the BioMagResBank (accession code: 19475).