Recent advances in segmental isotope labeling of proteins: NMR applications to large proteins and glycoproteins
Abstract In the last 15 years substantial advances have been made to place isotope labels in native and glycosylated proteins for NMR studies and structure determination. Key developments include segmental isotope labeling using Native Chemical Ligation, Expressed Protein Ligation and Protein Trans-Splicing. These advances are pushing the size limit of NMR spectroscopy further making larger proteins accessible for this technique. It is just emerging that segmental isotope labeling can be used to define inter-domain interactions in NMR structure determination. Labeling of post-translational modified proteins like glycoproteins remains difficult but some promising developments were recently achieved. Key achievements are segmental and site-specific labeling schemes that improve resonance assignment and structure determination of the glycan moiety. We adjusted the focus of this perspective article to concentrate on the NMR applications based on recent developments rather than on labeling methods themselves to illustrate the considerable potential for biomolecular NMR.
- Content Type Journal Article
- Pages 51-65
- DOI 10.1007/s10858-009-9362-7
- Authors
- Lenka Skrisovska, ETH Zürich Institute for Molecular Biology and Biophysics 8093 Zurich Switzerland
- Mario Schubert, ETH Zürich Institute for Molecular Biology and Biophysics 8093 Zurich Switzerland
- Frédéric H.-T. Allain, ETH Zürich Institute for Molecular Biology and Biophysics 8093 Zurich Switzerland
Source: Journal of Biomolecular NMR