Selenomethionine labeling of large biological macromolecular complexes: probing the structure of marine bacterial virus PM2.

Journal article

There is a need for improved tools for labeling protein species within large macromolecular assemblies. Here we describe a method for the efficient selenomethionine labeling of the membrane-containing bacterial virus PM2 for structural studies. By examining potential host cells a strain was found which was auxotrophic for methionine, and by performing a multiparameter search of conditions it was possible to derive a robust protocol which simultaneously minimized the toxic effects of the selenomethionine, so that a reasonable virus yield was maintained, whilst still achieving essentially complete labeling. This has allowed us to fingerprint the protein constituents of the virus in a relatively low resolution electron density map. Such a technique can be adapted to other macromolecule complexes studied by X-ray crystallography.

Authors: Kivelä, H; Abrescia, N; Bamford, J; Grimes, J; Stuart, D

• Publication status: Published
• Journal: Journal of structural biology
• Volume: 161
• Issue: 2
• Pages: 204-210
• Publication date: 2008-02-01
• DOI: 10.1016/j.jsb.2007.10.013
• EISSN: 1095-8657
• ISSN: 1047-8477
• Language: English
• Keywords: Crystallography, X-Ray; Corticoviridae; Methods; Selenomethionine