NMR experiments for resonance assignments of C-13, N-15 doubly-labeled flexible polypeptides: Application to the human prion protein hPrP(23-230) -- Web of Science version 4.1 (ISI2)

Easy Search Results--Full Record

Article 1 of 27

NMR experiments for resonance assignments of C-13, N-15 doubly-labeled flexible polypeptides: Application to the human prion protein hPrP(23-230)
Liu AZ, Riek R, Wider G, von Schroetter C, Zahn R, Wuthrich K
JOURNAL OF BIOMOLECULAR NMR 16: (2) 127-138 FEB 2000

Document type: Article

Language: English

Cited References: 46

Times Cited: 0

Abstract:
A combination of three heteronuclear three-dimensional NMR experiments tailored for sequential resonance assignments in uniformly N-15, C-13-labeled flexible polypeptide chains is described. The 3D (H)N(CO-TOCSY)NH, 3D (H)CA(CO-TOCSY)NH and 3D (H)CBCA(CO-TOCSY)NH schemes make use of the favorable N-15 chemical shift dispersion in unfolded polypeptides, exploit the slow transverse N-15 relaxation rates of unfolded polypeptides in high resolution constant-time [H-1, N-15]-correlation experiments, and use carbonyl carbon homonuclear isotropic mixing to transfer magnetization sequentially along the amino acid sequence. Practical applications are demonstrated with the 100-residue flexible tail of the recombinant human prion protein, making use of spectral resolution up to 0.6 Hz in the N-15 dimension, simultaneous correlation with the two adjacent amino acid residues to overcome problems associated with spectral overlap, and the potential of the presently described experiments to establish nearest-neighbor correlations across proline residues in the amino acid sequence.

Author Keywords:
carbonyl carbon homonuclear isotropic mixing, flexible polypeptide chains, human prion protein, sequential NMR assignment, triple-resonance experiments

KeyWords Plus:
ISOTOPICALLY ENRICHED PROTEINS, SIDE-CHAIN RESONANCES, BIOLOGICAL MACROMOLECULES, CORRELATION SPECTROSCOPY, IMPROVED SENSITIVITY, BACKBONE AMIDE, ENHANCEMENT, SPECTRA, PROTON, SEQUENCE

Addresses:
Wuthrich K, ETH Honggerberg, Inst Mol Biol & Biophys, CH-8093 Zurich, Switzerland.
ETH Honggerberg, Inst Mol Biol & Biophys, CH-8093 Zurich, Switzerland.

Publisher:
KLUWER ACADEMIC PUBL, DORDRECHT

IDS Number:
280YD

ISSN:
0925-2738

Article 1 of 27