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A pulsed field gradient isotope‐filtered 3D 13 C HMQC‐NOESY experiment for extracting intermolecular NOE contacts in molecular complexes
Author(s) -
Lee Woentae,
Revington Matthew J.,
Arrowsmith Cheryl,
Kay Lewis E.
Publication year - 1994
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(94)00740-3
Subject(s) - intermolecular force , two dimensional nuclear magnetic resonance spectroscopy , nuclear overhauser effect , chemistry , dephasing , isotope , analytical chemistry (journal) , crystallography , nuclear magnetic resonance , nuclear magnetic resonance spectroscopy , stereochemistry , molecule , physics , organic chemistry , chromatography , quantum mechanics
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.