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Rapid assignment of solution 31P NMR spectra of large proteins by solid-state spectroscopy.

Iuga A, Spoerner M, Ader C, Brunner E, Kalbitzer HR

Universität Regensburg, Institut für Biophysik und physikalische Biochemie, D-93040 Regensburg, Germany.

The application of the (31)P NMR spectroscopy to large proteins or protein complexes in solution is hampered by a relatively low intrinsic sensitivity coupled with large line widths. Therefore, the assignment of the phosphorus signals by two-dimensional NMR methods in solution is often extremely time consuming. In contrast, the quality of solid-state NMR spectra is not dependent on the molecular mass and the solubility of the protein. For the complex of Ras with the GTP-analogue GppCH(2)p we show solid-state (31)P NMR methods to be more sensitive by almost one order of magnitude than liquid-state NMR. Thus, solid-state NMR seems to be the method of choice for obtaining the resonance assignment of the phosphorus signals of protein complexes in solution. Experiments on Ras.GDP complexes show that the microcrystalline sample can be substituted by a precipitate of the sample and that unexpectedly the two structural states observed earlier in solution are present in crystals as well.

Published 19 June 2006 in Biochem Biophys Res Commun, 346(1): 301-5.
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