Solid-state 31P NMR spectroscopy of microcrystals of the Ras protein and its effector loop mutants: comparison between crystalline and solution state

Journal of Molecular Biology
Adriana IugaEike Brunner

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 (31)P 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 GppCH(2)p 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 o...Continue Reading

References

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Mar 17, 2004·Magnetic Resonance in Chemistry : MRC·Adriana Iuga, Eike Brunner

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Citations

Aug 4, 2010·Journal of Biomolecular NMR·Kumaran BaskaranHans Robert Kalbitzer
Jul 21, 2009·Journal of Magnetic Resonance·B HuJ P Amoureux
Jun 9, 2006·Biochemical and Biophysical Research Communications·Adriana IugaHans Robert Kalbitzer
Aug 3, 2013·Biochimica Et Biophysica Acta·Priyanka Prakash, Alemayehu A Gorfe
Feb 1, 2018·The Journal of Biological Chemistry·Daniel MannCarsten Kötting
Mar 27, 2015·Magnetic Resonance in Chemistry : MRC·Ga Hyang LeeYounkee Paik
Jul 5, 2005·The Journal of Biological Chemistry·Min YeTohru Kataoka
Oct 20, 2005·Magnetic Resonance in Chemistry : MRC·Michael SpoernerHans Robert Kalbitzer
Aug 21, 2013·Chembiochem : a European Journal of Chemical Biology·Abhishek Cukkemane, Marc Baldus
May 15, 2007·Biochemistry·Sandra M G Dias, Richard A Cerione

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