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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.

Original publication

DOI

10.1016/j.str.2007.01.011

Type

Journal article

Journal

Structure

Publication Date

03/2007

Volume

15

Pages

267 - 273

Keywords

Alkaloids, Cell Line, Cloning, Molecular, Enzyme Inhibitors, Genomics, Glycoproteins, Glycosylation, Humans, Swainsonine