Selenomethionine labeling of large biological macromolecular complexes: probing the structure of marine bacterial virus PM2.
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.
Kivelä
Hanna M
HM
Department of Biological and Environmental Science and Nanoscience Centre, University of Jyväskylä, P.O. Box 35 (Survontie 9), 40014 Jyväskylä, Finland.
Abrescia
Nicola G A
NG
Bamford
Jaana K H
JK
Grimes
Jonathan M
JM
Stuart
David I
DI
Bamford
Dennis H
DH
D019240:Corticoviridae / Q000737:chemistry* / Q000648:ultrastructure*; D018360:Crystallography, X-Ray; D008722:Methods; D012645:Selenomethionine / Q000737:chemistry*
D012645:Selenomethionine