Speeding Up Multidimensional NMR Data Acquisition

The power of NMR spectroscopy lies in its ability to obtain information about the structure and dynamics of individual atoms in complex molecules. The narrow frequency ranges of the NMR-active nuclei (₁H, ₁₃C, ₁₅N, ₃₁P) in biological macromolecules such as proteins, nucleic acids, and their complexes result in severe resonance overlap in one-dimensional spectra, making it practically impossible to extract atom-resolved information even for relatively small biopolymers. In order to deal with this problem, multidimensional NMR techniques are commonly used to spread and correlate the signals of individual nuclear spins along different frequency dimensions. A major drawback of conventional multidimensional NMR is the long experimental time needed for recording its data-a reflection of the hundreds or even thousands of experimental repetitions that are inherent to this kind of spectroscopy. While acquisition times for one-and two-dimensional spectra are on the order of seconds and minutes, respectively, typical three-dimensional NMR experimental times could be as long as days. For obtaining the four-and higher-dimensional spectra that may be of particular interest to the study of large molecular systems or of disordered proteins, acquisition times become altogether unreasonable. Moreover, as a large number of different spectra must be recorded during the NMR investigation of a macromolecule, this demands a high stability of the ensuing samples. Long experimental times also represent a major limitation for high-throughput studies as performed in the context of structural proteomics. It follows from all these considerations that reduced acquisition times could make data acquisition more efficient, and greatly enhance the structural and dynamic capabilities of biomolecular NMR. They could also enable real-time investigations of kinetic molecular processes, such as biochemical reactions, protein/RNA folding, and molecular assemblies by multidimensional spectroscopy. In this chapter, we review some of the fast multidimensional NMR data acquisition techniques that have emerged in recent years and their underlying concepts, and we discuss fields of biomolecular NMR research that could benefit from such fast NMR methods.