Posttranslational modifications of intact proteins detected by NMR spectroscopy: application to glycosylation.
Posttranslational modifications (PTMs) are an integral part of the majority of proteins. The characterization of structure and function of PTMs can be very challenging especially for glycans. Existing methods to analyze PTMs require complicated sample preparations and suffer from missing certain modifications, the inability to identify linkage types and thus chemical structure. We present a direct, robust, and simple NMR spectroscopy method for the detection and identification of PTMs in proteins. No isotope labeling is required, nor does the molecular weight of the studied protein limit the application. The method can directly detect modifications on intact proteins without sophisticated sample preparation. This approach is well suited for diagnostics of proteins derived from native organisms and for the quality control of biotechnologically produced therapeutic proteins.
Institute of Molecular Biology and Biophysics, ETH Zürich, 8093 Zürich (Switzerland). mario.schubert@sbg.ac.at.|Present address: Department of Molecular Biology, University of Salzburg, Billrothstrasse 11, 5020 Salzburg (Austria). mario.schubert@sbg.ac.at.
Mario
M
Schubert
Institute of Molecular Biology and Biophysics, ETH Zürich, 8093 Zürich (Switzerland). michal.walczak@mol.biol.ethz.ch.
Michal J
MJ
Walczak
Institute of Microbiology, ETH Zürich, 8093 Zürich (Switzerland).
Markus
M
Aebi
Institute of Molecular Biology and Biophysics, ETH Zürich, 8093 Zürich (Switzerland).
Gerhard
G
Wider
D002240:Carbohydrate Sequence; D006031:Glycosylation; D019906:Nuclear Magnetic Resonance, Biomolecular; D011134:Polysaccharides / Q000032:analysis / Q000737:chemistry; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism
D011134:Polysaccharides; D011506:Proteins
NMR spectroscopy; glycomics; glycoproteins; protein modifications; proteomics