Glycoprotein Structural Genomics: Solving the Glycosylation Problem
| Autor: | Joanne E. Nettleship, A R Aricescu, Simon J. Davis, Max Crispin, Kent S. Boles, Chack-Yung Yu, Raymond J. Owens, Raymond A. Dwek, E Y Jones, David I. Stuart, J A Fennelly, David Harvey, Veronica T. Chang, Edward J. Evans |
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| Rok vydání: | 2007 |
| Předmět: |
Glycosylation
Ways & Means Genomics Cell Line Structural genomics 03 medical and health sciences chemistry.chemical_compound Endoglycosidase H Alkaloids Structural Biology Humans Cloning Molecular Enzyme Inhibitors Molecular Biology Glycoproteins 030304 developmental biology chemistry.chemical_classification 0303 health sciences biology Swainsonine 030302 biochemistry & molecular biology Enzyme Kifunensine chemistry Biochemistry Cell culture biology.protein Glycoprotein |
| Zdroj: | Structure(London, England:1993) |
| ISSN: | 0969-2126 |
| DOI: | 10.1016/j.str.2007.01.011 |
| Popis: | Summary Glycoproteins present special problems for structural genomic analysis because they often require glycosylation in order to fold correctly, whereas their chemical and conformational heterogeneity generally inhibits crystallization. We show that the “glycosylation problem” can be solved by expressing glycoproteins transiently in mammalian cells in the presence of the N-glycosylation processing inhibitors, kifunensine or swainsonine. This allows the correct folding of the glycoproteins, but leaves them sensitive to enzymes, such as endoglycosidase H, that reduce the N-glycans to single residues, enhancing crystallization. Since the scalability of transient mammalian expression is now comparable to that of bacterial systems, this approach should relieve one of the major bottlenecks in structural genomic analysis. |
| Databáze: | OpenAIRE |
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