Eukaryotic expression:Developments for structural proteomics

Aricescu, A.R., Assenberg, R., Bill, Roslyn M., Busso, D., Chang, V.T., Davis, S.J., Dubrovsky, A., Gustafsson, L., Hedfalk, K., Heinemann, U., Jones, I.M., Ksiazek, D., Lang, C., Maskos, K., Messerschmidt, A., Macieira, S., Peleg, Y., Perrakis, A., Poterszman, A., Schneider, G., Sixma, T.K., Sussman, J.L., Sutton, G., Tarboureich, N., Zeev-Ben-Mordehai, T. and Jones, E. Yvonne (2006). Eukaryotic expression:Developments for structural proteomics. Acta Crystallographica: Section D, 62 (10), pp. 1114-1124.

Abstract

The production of sufficient quantities of protein is an essential prelude to a structure determination, but for many viral and human proteins this cannot be achieved using prokaryotic expression systems. Groups in the Structural Proteomics In Europe (SPINE) consortium have developed and implemented high-throughput (HTP) methodologies for cloning, expression screening and protein production in eukaryotic systems. Studies focused on three systems: yeast (Pichia pastoris and Saccharomyces cerevisiae), baculovirus-infected insect cells and transient expression in mammalian cells. Suitable vectors for HTP cloning are described and results from their use in expression screening and protein-production pipelines are reported. Strategies for co-expression, selenomethionine labelling (in all three eukaryotic systems) and control of glycosylation (for secreted proteins in mammalian cells) are assessed. © International Union of Crystallography, 2006.

Publication DOI: https://doi.org/10.1107/S0907444906029805
Divisions: Life & Health Sciences > Biosciences
Life & Health Sciences
Life & Health Sciences > Cellular and Molecular Biomedicine
Additional Information: Copyright © International Union of Crystallography Author(s) of this paper may load this reprint on their own web site or institutional repository provided that this cover page is retained. Republication of this article or its storage in electronic databases other than as specified above is not permitted without prior permission in writing from the IUCr.
Uncontrolled Keywords: baculovirus,eukaryotic expression,high throughput,mammalian cells,yeast,Clinical Biochemistry,Biochemistry, Genetics and Molecular Biology(all),Biochemistry,Biophysics,Condensed Matter Physics,Structural Biology
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Published Date: 2006-10
Authors: Aricescu, A.R.
Assenberg, R.
Bill, Roslyn M. ( 0000-0003-1331-0852)
Busso, D.
Chang, V.T.
Davis, S.J.
Dubrovsky, A.
Gustafsson, L.
Hedfalk, K.
Heinemann, U.
Jones, I.M.
Ksiazek, D.
Lang, C.
Maskos, K.
Messerschmidt, A.
Macieira, S.
Peleg, Y.
Perrakis, A.
Poterszman, A.
Schneider, G.
Sixma, T.K.
Sussman, J.L.
Sutton, G.
Tarboureich, N.
Zeev-Ben-Mordehai, T.
Jones, E. Yvonne

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