Protein 3D structure determination by high-resolution solid-state NMR

摘要

Developments in NMR technology, sample preparation, pulse sequence methodology and structure calculation protocols have recently allowed one to progress towards structure determination at high-resolution of proteins by solid-state NMR spectroscopy. We here report solid-state NMR protocols based on magic-angle-spinning experiments, combined with modified structure calculation protocols, for structure determination of uniformly ~(13)C, ~(15)N isotopically labeled proteins. We demonstrate the use of these protocols to obtain high-resolution structures for the example of the microcrystalline Crh protein. The CHHC, DARR and PAR solid-state NMR experiments, as well as the calculation protocols using the program ARIA, are presented.