Two-dimensional chemical exchange and cross-relaxation spectroscopy of coupled nuclear spins

Macura, Slobodan; Huang, Yihui; Suter, Dieter; Ernst, R. R.

doi:10.1016/0022-2364(81)90037-8

Cited by 635 publications

(530 citation statements)

References 17 publications

Supporting

Mentioning

526

Contrasting

Order By: Relevance

“…Spectral widths were 14.12 ppm in both dimensions. NOESY (Macura et al, 1981), DQF-COSY (Rance et al, 1983), and clean-TOCSY experiments (Griesinger et al, 1988) were performed in both H 2 0 and DzO. Two-dimensional NOESY spectra were recorded with a mixing time of 100 ms, during which the solvent was irradiated.…”

Section: 'H"h 2 0 Nmrmentioning

confidence: 99%

Solution structure of the DNA‐binding domain of the heat shock transcription factor determined by multidimensional heteronuclear magnetic resonance spectroscopy

et al. 1994

View full text Add to dashboard Cite

The solution structure of the 92-residue DNA-binding domain of the heat shock transcription factor from Kluyveromyces fuctis has been determined using multidimensional NMR methods. Three-dimensional (3D) triple resonance, 'H-'3C-'3C-'H total correlation spectroscopy, and I5N-separated total correlation spectroscopyheteronuclear multiple quantum correlation experiments were used along with various 2D spectra to make nearly complete assignments for the backbone and side-chain 'H, "N, and I3C resonances. Five-hundred eighty-three NOE constraints identified in 3D I3C-and "N-separated NOE spectroscopy (N0ESY)-heteronuclear multiple quantum correlation spectra and a 4-dimensional I3C/l3C-edited NOESY spectrum, along with 35 6, 9 xI , 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 P-sheet. The backbone RMS deviation (RMSD) for the family of structures was 1.03 * 0.19 A with respect to the average structure. The topology is analogous to that of the C-terminal domain of the catabolite gene activator protein and appears to be in the helix-turn-helix family of DNAbinding proteins. The overall fold determined by the NMR data is consistent with recent crystallographic work on this domain (Harrison CJ, Bohm AA, Nelson HCM, 1994, Science 263:224) as evidenced by RMSD between backbone atoms in the NMR and X-ray structures of 1.77 k 0.20 A. Several differences were identified some of which may be due to protein-protein interactions in the crystal.

show abstract

Section: 'H"h 2 0 Nmrmentioning

confidence: 99%

Solution structure of the DNA‐binding domain of the heat shock transcription factor determined by multidimensional heteronuclear magnetic resonance spectroscopy

et al. 1994

View full text Add to dashboard Cite

show abstract

“…The TOCSY experiments (Braunschweiler & Ernst, 1983) were recorded using either the MLEV-17 (Bax & Davis, 1985) or DIPSI-2 (Rucker & Shaka, 1989) spin lock sequences, with a mixing time of 41 ms. The NOESY spectra (Macura et al, 1981) were recorded with mixing times of 120-160 ms. Typically, 400-500tI increments were acquired, with 128-256 scans per increment.…”

Section: Nmr Spectroscopymentioning

confidence: 99%

Role of the C‐terminus in the activity, conformation, and stability of interleukin‐6

et al. 1993

View full text Add to dashboard Cite

Two murine interleukin-6 (mIL-6) variants were constructed using the polymerase chain reaction (PCR), one lacking the last five residues (183-187) at the C-terminus (pMC5) and another with the last five residues of mIL-6 substituted by the corresponding residues of human IL-6 (pMC5H). The growth stimulatory activity of pMC5 on the mouse hybridoma cell line 7TD1 was <0.05% of mIL-6, whereas pMC5H and mIL-6 were equipotent. The loss of biological activity of pMCS correlated with its negligible receptor binding affinity on 7TD1 cells, while the binding of pMC5H was comparable to that of mIL-6. Both pMC5 and pMCSH, like mIL-6, failed to interact with recombinant soluble human IL-6 receptor when assayed by surface plasmon resonance-based biosensor analysis. These studies suggest that the C-terminal seven amino acids of human IL-6, alone, do not define species specificity for receptor binding.A variety of biophysical techniques, as well as the binding of a conformational-specific monoclonal antibody, indicated that the global fold of the mIL-6 variants was similar to that of mIL-6, although small changes in the NMR spectra, particularly for pMC5, were observed. Some of these changes involved residues widely separated in the primary structure. For instance, interactions involving Tyr-22 were influenced by the C-terminal amino acids suggesting that the N-and C-termini of mIL-6 are in close proximity. Equilibrium unfolding experiments indicated that pMC5 was 0.8 kcal/mol less stable than mIL-6, whereas pMCSH was 1.4 kcal/mol more stable. These studies emphasize the structural importance of the C-terminal amino acids of IL-6 and suggest that truncation or mutation of this region could lead to small but significant alterations in other regions of the molecule.

show abstract

“…Based on the solution structure of Ets-lAN331, there are distinct regions of negative and positive charges on the surfaces of these proteins that may result in the need for high salt concentrations to screen intermolecular electrostatic interactions (Donaldson et al, 1996). 'H-'H 2 0 NMR spectroscopy 2D NOESY (Macura et al, 1981) and clean-TOCSY (Griesinger et al, 1988) experiments were recorded on unlabeled Ets-IAN280 in 99% 'H20. Low-power continuous radiofrequency irradiation with the carrier placed on the water resonance (4.70 ppm at 32 "C) was used to suppress the residual water signal.…”

Section: Solution Conditionsmentioning

confidence: 99%

Structural coupling of the inhibitory regions flanking the ETS domain of murine Ets‐1

et al. 1996

View full text Add to dashboard Cite

Several members of the ets gene family of transcription factors show negative regulation of DNA binding by intramolecular interactions. A structural mechanism for this auto-inhibition is investigated using a 161-residue N-terminal deletion mutant of murine Ets-1, Ets-1AN280. This protein shows a similar reduced affinity for DNA as native Ets-1 because it contains the ETS domain in context of the flanking amino-and carboxy-terminal regions that together mediate repression of DNA binding. The secondary structure of Ets-IAN280 was determined using NMR chemical shift, NOE, J coupling, and amide hydrogen exchange information. In addition to the winged helix-turn-helix ETS domain, Ets-1AN280 contains two a-helices in the amino-terminal inhibitory region and one a-helix in the carboxy-terminal inhibitory region. Chemical shift comparisons were made between this protein and an activated form of Ets-1 lacking the amino-terminal inhibitory region. The spectral differences demonstrate that the amino-and carboxy-terminal inhibitory sequences are structurally coupled to one another, thus explaining the observation that both regions are required for the repression of DNA binding. Furthermore, these data show that the inhibitory sequences also interact directly with the first helix of the intervening ETS domain, thereby providing a pathway for the repression of DNA binding. These results lead to a model of an inhibitory module in Ets-1 composed of both the amino-and carboxy-terminal regions interfaced with the ETS domain. This establishes the structural framework for understanding the intramolecular inhibition of Ets-1 DNA binding.Keywords: allosteric; DNA binding; ETS domain; Ets-1 and Ets-2; intramolecular inhibition; NMR; protein structure; winged helix-turn-helix DNA binding, activation, and oligomerization associated with a particular transcription factor may be governed by its subcellular localization, posttranslational modifications, ligand binding, and protein-protein interactions.Recently, data have Abbreviations: ets, E26 transformation specific; Ets-1AC428, the carboxy-terminal deletion fragment of murine Ets-1 consisting of residues 1-428; Ets-1AN280, the amino-terminal deletion fragment of murine Ets-1 consisting of residues 280-440; Ets-1AN331, the aminoterminal deletion fragment of murine Ets-1 consisting of residues 33 1-440; FID, free induction decay; HSQC, heteronuclear single quantum correlation; NOESY, nuclear Overhauser effect spectroscopy; TOCSY, total correlation spectroscopy; IPTG, isopropyl-8-D-thiogalactopyranoside; DTT, dithiothreitol; pH*, the observed pH meter reading without correction for isotope effects; wHTH, winged helix-turn-helix; HSF, heat shock transcription factors; DSS, 2,2-dimethyl-2-silapentane-5-sulfonate, sodium salt.

show abstract

Two-dimensional chemical exchange and cross-relaxation spectroscopy of coupled nuclear spins

Cited by 635 publications

References 17 publications

Solution structure of the DNA‐binding domain of the heat shock transcription factor determined by multidimensional heteronuclear magnetic resonance spectroscopy

Solution structure of the DNA‐binding domain of the heat shock transcription factor determined by multidimensional heteronuclear magnetic resonance spectroscopy

Role of the C‐terminus in the activity, conformation, and stability of interleukin‐6

Structural coupling of the inhibitory regions flanking the ETS domain of murine Ets‐1

Contact Info

Product

Resources

About