Document Detail

NMR structure determination of a segmentally labeled glycoprotein using in vitro glycosylation.
MedLine Citation:
PMID:  19154179     Owner:  NLM     Status:  MEDLINE    
Although there is great interest in three-dimensional structures of glycoproteins and complex oligosaccharides, their structural determination have been hampered by inhomogeneous and incomplete glycosylation, poor expression, low tendency to crystallize, and severe chemical shift overlap. Using segmental labeling of the glycan and the protein component by in vitro glycosylation, we developed a novel method of NMR structural determination that overcomes some of these problems. Highly homogeneously glycosylated proteins in milligram amounts can be obtained. This allowed the determination of the structure of an N-linked glycoprotein from Campylobacter jejuni. The glycosylation acceptor site was found to be in a flexible loop. The presented methodology extends the observable NOE distance limit of oligosaccharides significantly over 4 A, resulting in a high number of distance restraints per glycosidic linkage. A well-defined glycan structure was obtained.
Vadim Slynko; Mario Schubert; Shin Numao; Michael Kowarik; Markus Aebi; Frédéric H-T Allain
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of the American Chemical Society     Volume:  131     ISSN:  1520-5126     ISO Abbreviation:  J. Am. Chem. Soc.     Publication Date:  2009 Jan 
Date Detail:
Created Date:  2009-01-21     Completed Date:  2009-03-10     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7503056     Medline TA:  J Am Chem Soc     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1274-81     Citation Subset:  IM    
Institute of Molecular Biology and Biophysics, ETH Zurich, CH-8093 Zurich, Switzerland.
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MeSH Terms
Amino Acid Sequence
Campylobacter jejuni / chemistry
Glycoproteins / analysis*,  chemistry*
Magnetic Resonance Spectroscopy / methods*
Models, Molecular
Protein Structure, Tertiary
Reg. No./Substance:

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