Abstract
The described TROSY-based experiments for investigating backbone dynamics of proteins make it possible to elucidate internal motions in large proteins via measurements of T 1 , T 2 , and NOE of backbone 15 N nuclei. In our proposed sequences, the INEPT sequence is eliminated and the PEP sequence is replaced by the ST2-PT sequence from the HSQC-based experiments. This has the benefit of shortening the pulse sequences by 5.4 ms (=1/2J) and results in an increase in the intrinsic sensitivity of the proposed TROSY-based experiments. The TROSY-based experiments are on average of 13% more sensitive than the corresponding HSQC-based experiments on a uniformly 15 N-labeled Xenopus laevis calcium-bound calmodulin sample on a 750-MHz spectrometer at 5°C. The amide proton linewidths of the TROSY-based experiments are 2-13 Hz narrower than those of the HSQC experiments. More sensitivity gain and higher resolution are expected if the protein sample is deuterated.
Original language | English (US) |
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Pages (from-to) | 423-426 |
Number of pages | 4 |
Journal | Journal of Magnetic Resonance |
Volume | 143 |
Issue number | 2 |
DOIs | |
State | Published - Apr 2000 |
Bibliographical note
Funding Information:This work is supported by grants from the Research Grant Council of Hong Kong (HKUST6197/97M, HKUST6038/98M, and HKUST6199/99M). The Hong Kong Biotechnology Research Institute is acknowledged for the purchase of the 750-MHz NMR spectrometer. LKN acknowledges support from the National Science Foundation (MCB-9808727) and the National Institutes of Health (1 R01 CA77478-01).
Keywords
- NMR
- Protein dynamics
- Relaxation
- TROSY