Stumber, Michael and Geyer, Matthias and Graf, Robert and Kalbitzer, Hans Robert and Scheffzek, Klaus and Haeberlen, Ulrich (2002) Observation of slow dynamic exchange processes in Ras protein crystals by P-31 solid state NMR spectroscopy. JOURNAL OF MOLECULAR BIOLOGY, 323 (5). pp. 899-907. ISSN 0022-2836, 1089-8638
Full text not available from this repository. (Request a copy)Abstract
The folding, structure and biological function of many proteins are inherently dynamic properties of the protein molecule. Often, the respective molecular processes are preserved upon protein crystallization, leading, in X-ray diffraction experiments, to a blurring of the electron density map and reducing the resolution of the derived structure. Nuclear magnetic resonance (NMR) is known to be an alternative method to study molecular structure and dynamics. We designed and built a probe for phosphorus solid state NMR that allows for the first time to study static properties as well as dynamic processes in single-crystals of a protein by NMR spectroscopy. The sensitivity achieved is sufficient to detect the NMR signal from individual phosphorus sites in a 0.3 mm(3) size single-crystal of GTPase Ras bound to the nucleotide GppNHp, that is, the signal from approximately 10(15) phosphorus nuclei. The NMR spectra obtained are discussed in terms of the conformational variability of the active center of the Ras-nucleotide complex. We conclude that, in the crystal, the protein complex exists in three different conformations. Magic angle spinning (MAS) NMR spectra of a powder sample of Ras-GppNHp show a splitting of one of the phosphate resonances and thus confirm this conclusion. The MAS spectra provide, furthermore, evidence of a slow, temperature-dependent dynamic exchange process in the Ras protein crystal. (C) 2002 Elsevier Science Ltd. All rights reserved.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | MOLECULAR SWITCH; GTP HYDROLYSIS; SIGNAL-TRANSDUCTION; COMPLEX; P21; BINDING; CONFORMATION; RESOLUTION; EFFECTORS; P21(RAS); solid state NMR; MAS NMR; protein dynamics; dynamics in crystals; Ras |
| Subjects: | 500 Science > 530 Physics 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie > Prof. Dr. Dr. Hans Robert Kalbitzer |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 26 Aug 2021 12:32 |
| Last Modified: | 26 Aug 2021 12:32 |
| URI: | https://pred.uni-regensburg.de/id/eprint/39675 |
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