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A pulsed field gradient isotope‐filtered 3D 13C HMQC‐NOESY experiment for extracting intermolecular NOE contacts in molecular complexes
Abstract
A pulsed field gradient three‐dimensional isotope‐filtered 13 C HMQC‐NOESY experiment has been developed to characterize intermolecular contacts in a 37 kDa macromolecular ternary complex consisting of uniformly 13 C labeled trp‐repressor, its natural abundance co‐repressor, l‐tryptophan, and natural abundance operator DNA. The pulse scheme makes use of pulsed field gradients for the removal of artifacts and dephasing of unwanted magnetization during isotope filtering, and employs a strategy to minimize the time that magnetization resides in the transverse plane. The experiment provides solely intermolecular NOE contacts between protons of the labeled protein and protons of the unlabeled species, and has proven to be especially useful in eliminating ambiguities between intra‐ and intermolecular NOEs in the isotope‐edited 3D 13 C HMQC‐NOESY spectrum of the complex.
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Cited by
Fourmy, D., Recht, M. I., Blanchard, S. C., & Puglisi, J. D. (1996) · Science · doi:10.1126/science.274.5291.1367 · Cited by 829
Radhakrishnan, I., Pérez-Alvarado, G. C., Parker, D., Dyson, H. J., Montminy, M. R., & Wright, P. E. (1997) · Cell · doi:10.1016/s0092-8674(00)80463-8 · Cited by 728
Structure of the HIV-1 Nucleocapsid Protein Bound to the SL3 Ψ-RNA Recognition Element
De Guzman, R. N., Wu, Z. R., Stalling, C. C., Pappalardo, L., Borer, P. N., & Summers, M. F. (1998) · Science · doi:10.1126/science.279.5349.384 · Cited by 659
Byeon, I.-J. L., Meng, X., Jung, J., Zhao, G., Yang, R., Ahn, J., Shi, J., Concel, J., Aiken, C., Zhang, P., et al. (2009) · Cell · doi:10.1016/j.cell.2009.10.010 · Cited by 253
Metadata
- Authors
- Lee, W., Revington, M. J., Arrowsmith, C., & Kay, L. E.
- Venue
- FEBS Letters
- Publisher
- Wiley
- Published
- 1994-08-15
- Volume
- 350
- Issue
- 1
- Pages
- 87-90
- ISSN
- 0014-5793 (ISSN-L)
- 1873-3468
- OpenAlex
- W1980064805
- Cited by
- 162
- References
- 0
- FWCI
- 4.76
- Type
- journal article
- Language
- en
- Open access
- closed
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- Lee, W., Revington, M. J., Arrowsmith, C., & Kay, L. E. (1994). A pulsed field gradient isotope‐filtered 3D 13C HMQC‐NOESY experiment for extracting intermolecular NOE contacts in molecular complexes. FEBS Letters. https://doi.org/10.1016/0014-5793(94)00740-3
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