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3D HMQC-NOESY-HMQC
DESCRIPTION
The 3D HMQC-NOESY-HMQC experiment is specifically designed to obtain X-edited NOESY spectra of X-labeled biomolecules and protein-ligand complexes from which homonuclear 1H-1H NOEs can be clearly assigned even in overcrowded regions and even between protons with degenerate chemical shifts. In fact, this pulse sequence is derived from a 4D HMQC-NOESY-HMQC experiment in which one variable period has been fixed or from a 2D w1,w2-edited NOESY experiment (See correlation between NOE experiments). Similar results can be obtained from analogs 3D HSQC-NOESY-HSQC experiment.
REQUIREMENTS
Implementation on AVANCE spectrometers is feasible. Improved versions using pulsed field gradients (PFGs) are also available and, therefore, in such cases gradient technology is required. The 1H-13C HMQC-NOESY-HMQC experiment is usually applied on 13C-labeled proteins in D20 and the 1H-15N HMQC-NOESY-HMQC is usually applied on 15N-labeled molecules in H20.
VERSIONS
The original 3D HMQC-NOESY-HMQC pulse sequence ( 90JACS9020 and 97BIO3959 ) basically consists of three parts:
  • A 2D 1H-15N HMQC pulse train
  • A fixed 1H-1H NOESY mixing time
  • A second 2D 1H-15N HMQC pulse train

    • Several improved and modified versions have been reported:

  • HMQC pulse trains are replaced by analogs HSQC pulse trains, rendering HMQC-NOESY-HSQC ( 94BIO14858 ) or HSQC-NOESY-HSQC ( 95JMRB294-108 and 99JB177-15 ) pulse sequences.
  • Analogs 15N-HSQC-NOESY-15N-TROSY and 15N-TROSY-NOESY-15N-TROSY experiments have been also reported ( 00JB261-18 ).
  • The first step is a 1H-13C transfer magnetization ( 99JB177-15 ) giving a 1H-13C-HSQC-NOESY- 1H-15N-HSQC experiment
  • The last step is a 1H-13C transfer magnetization ( 99JB177-15 ) giving a 1H-15N-HSQC-NOESY- 1H-13C-HSQC experiment
  • Time-shared 15N,13C-separated NOESY experiments in which 13C and 15N labeling can be encoded in separate time domains (SeeTime-shared simultaneous acquisition)
    • EXPERIMENTAL DETAILS
    More details on practical implementation of the 3D HMQC-NOESY-HMQC experiment on AVANCE spectrometers can be found in the corresponding Tutorial 3D HMQC-NOESY-HMQC experiment
    SPECTRA
    The 1H-X-HMQC-NOESY-1H-Y-HMQC experiment affords a 3D spectrum in which 1H(i), X(i) and Y(j) chemical shifts are displayed in three independent dimensions.
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