In this work we present a new pulse sequence for the measurement of long-range heteronuclear coupling constants in which the optimization of coherence selection by pulsed field gradients offers a net increase in sensitivity. This type of experiments is extremely valuable for conformational studies o
Sensitivity Gain by Simultaneous Acquisition of Two Coherence Pathways: The HNCA+ Experiment
โ Scribed by Michael Salzmann; Alfred Ross; Michael Czisch; Gerhard Wider
- Publisher
- Elsevier Science
- Year
- 2000
- Tongue
- English
- Weight
- 96 KB
- Volume
- 143
- Category
- Article
- ISSN
- 1090-7807
No coin nor oath required. For personal study only.
โฆ Synopsis
In most multidimensional nuclear magnetic resonance experiments a single and distinct coherence transfer pathway is selected by phase cycling or by pulsed field gradients. It was shown that simultaneously exploiting more than one coherence transfer pathway could increase the overall sensitivity of NMR experiments. However, sensitivity enhancement schemes described to date introduce additional delays in the pulse schemes, resulting in considerable decrease of the expected sensitivity gain when applied to biomolecules due their fast transverse relaxation. A novel sensitivity enhancement principle which increases sensitivity of an experiment by simultaneously exploiting two completely independent coherence pathways in a single NMR pulse scheme is presented in this paper. As an example an improved HNCA experiment, the HNCA(+), is presented, which combines the "out-and-back" coherence transfer pathway used in HNCA with an "out-and-stay" experiment, analogous to HCANH, without adding any time periods compared to the conventional HNCA pulse sequence. The applicability of the HNCA(+) was theoretically evaluated with regard to different sizes of peptides or proteins, which showed that the experimental time can be reduced twofold in ideal cases. The application of this novel experiment to a 7-kDa protein showed a 20% sensitivity gain of HNCA(+) when compared to conventional HNCA.
๐ SIMILAR VOLUMES