Biomedical Engineering Reference
In-Depth Information
eukaryotic, and preferably mammalian, cells. While the seminal work of
Selenko
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demonstrated that in-cell NMR in eukaryotic cells was feasible, this
work relied on injection of purified, isotope-labelled protein into Xenopus
oocytes. More recently the elegant work of Inomata and colleagues has
demonstrated high-resolution HSQC spectra of proteins inside living human
cells. This method uses cell penetrating peptides
69
to deliver protein into the
cytosol. The peptides are covalently linked to the protein and the release is
made upon enzymatic activity or reductive cleavage.
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Importantly, the
method was used to show both FK506 and rapamycin binding to
15
N-labelled
FKBP in HeLa cells.
11.6 Perspectives
In 2008, a group of experienced NMR spectroscopists in industry and
academia wrote on the perspective for the use of NMR for drug discovery.
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It
is greatly exciting to note that many of the developments that have been
highlighted in this review are in areas that the authors of the perspective
defined as being important for the future expansion of NMR within industry.
Although NMR now faces increased competition from other methods in the
area of ligand discovery, the value of its contributions throughout the drug-
discovery pipeline make the investment in instrumentation and expertise well
worthwhile.
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