Solution structure of the DNA-binding domain of the heat shock transcription factor determined by multidimensional heteronuclear magnetic resonance spectroscopy
- Univ. of California, Berkeley, CA (United States)
- Lawrence Berkeley Lab., CA (United States). Structural Biology Division
The solution structure of the 92-residue DNA-binding domain of the heat shock transcription factor from Kluyveromyces lactis has been determined using multidimensional NMR methods. Three-dimensional (3D) triple resonance, {sup 1}H-{sup 13}C-{sup 13}C-{sup 1}H total correlation spectroscopy, and {sup 15}N-separated total correlation spectroscopy-heteronuclear multiple quantum correlation experiments were used along with various 2D spectra to make nearly complete assignments for the backbone and side-chain {sup 1}H, {sup 15}N, and {sup 13}C resonances. Five-hundred eighty-three NOE constraints identified in 3D {sup 13}C- and {sup 15}N-separated NOE spectroscopy (NOESY)-heteronuclear multiple quantum correlation spectra and a 4-dimensional {sup 13}C/{sup 13}C-edited NOESY spectrum, along with 35 {phi}, 9{chi}{sub 1}, and 30 hydrogen bond constraints, were used to calculate 30 structures by a hybrid distance geometry/simulated annealing protocol, of which 24 were used for structural comparison. The calculations revealed that a 3-helix bundle packs against a small 4-stranded antiparallel {beta}-sheet. The backbones RMS deviation (RMSD) for the family of structures was 1.03{+-}0.19 {angstrom} with respect to the average structure. The topology is analogous to that of the C-terminal domain of the catabolite gene activator protein and papers to be in the helix-turn-helix family of DNA-binding proteins. The overall fold determined by the NMR data is consistent with recent crystallographic work on this domain as evidenced by RMSD between backbone atoms in the NMR and X-ray structures of 1.77{+-}0.20{angstrom}. Several differences were identified some of which may be due to protein-protein interactions in the crystal.
- Sponsoring Organization:
- USDOE
- OSTI ID:
- 136718
- Journal Information:
- Protein Science, Vol. 3; Other Information: PBD: 1994
- Country of Publication:
- United States
- Language:
- English
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