Progress in Nuclear Magnetic Resonance Spectroscopy. Practical aspects of NMR signal assignment in larger and challenging proteins

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Progress in Nuclear Magnetic Resonance Spectroscopy. Practical aspects of NMR signal assignment in larger and challenging proteins

Publication date: Available online 15 December 2013
Source:Progress in Nuclear Magnetic Resonance Spectroscopy

Author(s): Dominique P Frueh

NMR has matured into a technique routinely employed for studying proteins in near physiological conditions. However, applications to larger proteins are impeded by the complexity of the various correlation maps necessary to assign NMR signals. This article reviews the data analysis techniques traditionally employed for resonance assignment and describes alternative protocols necessary for overcoming challenges in large protein spectra. In particular, simultaneous analysis of multiple spectra may help overcome ambiguities or may reveal correlations in an indirect manner. Similarly, visualization of orthogonal planes in a multidimensional spectrum can provide alternative assignment procedures. We describe examples of such strategies for assignment of backbone, methyl, and nOe resonances. We describe experimental aspects of data acquisition for the related experiments and provide guidelines for preliminary studies. Focus is placed on large folded monomeric proteins and examples are provided for 37, 48, 53, and 81 kDa proteins.
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