PROTON-DETECTED HETERONUCLEAR NMR SPECTROSCOPY

Project: Research project

Project Details

Description

We wish to modify extensively a commercial NMR spectrometer to provide an
exceptionally high sensitivity proton-detected NMR spectroscopy of lower
frequency nuclei, (abbreviated for convenience to HUMP, Heteronuclear
Multiquantum Proton Detected NMR) particularly for 15N, 13C, and Cd, using
multiquantum evolution methods in two-dimensional NMR. Modifications of
frequency generation, pulse programming, and probes will be required. For
15N, we have demonstrated a three order of magnitude increase in
sensitivity compared to current direct detection, and hope to obtain an
additional two orders of magnitude by use of state-of-the-art electronics
and improved probe designs. This will provide a completely new area of
research in biomolecular structure investigation of proteins and nucleic
acids, using materials with natural abundance 15N content. With 15N
labeled material, HUMP can provide an unprecedently selective and sensitive
probe NMR method for site specific investigation of enzyme mechanisms or
effector-receptor interactions. Applications to nuclei other than 15N can
potentially provide sensitivity gains, which, while lower, are nonetheless
significant in combination with the two-dimensional resolution of HUMP.
The development of improved indirect detection techniques will also be of
general application in studies of metabolism using stable isotope labeled
materials.

The three year period of this proposal involves a 24-month implementation
period, followed by a 12-month period of use of the instrument, on projects
to which this specialized instrument can make unique contributions.
StatusFinished
Effective start/end date1/1/901/1/90

Funding

  • National Institute of General Medical Sciences

ASJC

  • Analytical Chemistry
  • Nuclear and High Energy Physics
  • Molecular Biology
  • Structural Biology
  • Biophysics

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