Iuga, Adriana and Spoerner, Michael and Kalbitzer, Hans Robert and Brunner, Eike (2004) Solid-state P-31 NMR spectroscopy of microcrystals of the Ras protein and its effector loop mutants: Comparison between crystalline and solution state. JOURNAL OF MOLECULAR BIOLOGY, 342 (3). pp. 1033-1040. ISSN 0022-2836, 1089-8638
Full text not available from this repository. (Request a copy)Abstract
Cycling between a GTP bound "on" state and a GDP bound "off" state, guanine nucleotide-binding (GNB) proteins act as molecular switches. The switching process and the interaction with effectors, GTPase-activating proteins, and guanosine nucleotide-exchange factors is accompanied by pronounced conformational changes of the switch regions of the GNB proteins. The aim of the present contribution is to correlate conformational changes observed by liquid-state NMR with solid-state P-31 NMR data and with the results of X-ray crystallography. Crystalline wild-type Ras complexed with GTP analogs such as GppCH(2)p and GppNHp could be prepared. At low temperatures, two different signals were found for the gamma-phosphate group of GppNHp bound to wild-type Ras. This behavior indicates the existence of two different conformations of the molecule in the crystalline state as it is found in solution but not by X-ray crystallography. In contrast to the GppNHp complex, the two separate gamma-phosphate signals could not be observed for GppCH2p bound to wild-type Ras. However, an increasing linewidth at low temperature indicates the presence of an exchange process. The results obtained for the wild-type protein are compared with the behavior of GppNHp complexes of the effector loop mutants Ras(T35S) and Ras(T35A). These mutants prefer a conformation similar to the GDP bound "off" state. (C) 2004 Elsevier Ltd. All rights reserved.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | PHOSPHORYLATED AMINO-ACIDS; ROTATING SOLIDS; CHEMICAL-SHIFTS; P21; PHOSPHATES; RESOLUTION; RESONANCE; EXCHANGE; SYSTEMS; SPECTRA; Ras protein; effector loop mutants; molecular switches; phosphorous solid-state NMR spectroscopy |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie > Alumni or Retired > Prof. Dr. Eike Brunner Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie > Prof. Dr. Dr. Hans Robert Kalbitzer |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 28 Jun 2021 11:47 |
| Last Modified: | 28 Jun 2021 11:47 |
| URI: | https://pred.uni-regensburg.de/id/eprint/37186 |
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