In-Cell NMR

Synonyms

NMR

In-cell NMR is a multidimensional NMR spectroscopy of macromolecules such as proteins and nucleic acids in living cells, aiming to analyze their structures and functions. Techniques of isotope-aided solution NMR spectroscopy are used to selectively derive signals of macromolecules of interest from those of a large number of other molecules present in cells.

So far, E. coli cells, Xenopus laevis oocytes, human HeLa cells, and monkey COS7 and 293F cells have been used as “host cells” for the in-cell approach. For E. coli cells, proteins to be analyzed are over-expressed in cells grown in isotope-enriched media, so that the proteins are semi-selectively labeled with NMR-active nuclei such as ¹⁵N and ¹³C. The whole cells are subjected to NMR measurements, and appropriate pulse schemes are employed to allow only the signals of labeled molecules to be detected. The obtained spectra possibly suffer from NMR signals of intrinsic E. coliproteins and other endogenous molecules...