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).