Journal
PROTEIN SCIENCE
Volume 18, Issue 5, Pages 1121-1127Publisher
WILEY
DOI: 10.1002/pro.97
Keywords
X-ray crystallography; selenomethionine incorporation; Lactococcus lactis; mass spectrometry
Categories
Ask authors/readers for more resources
Lactococcus lactis is a promising host for ( membrane) protein overproduction. Here, we describe a protocol for incorporation of selenomethionine ( SeMet) into proteins expressed in L. lactis. Incorporation efficiencies of SeMet in the membrane protein complex OpuA (an ABC transporter) and the soluble protein OppA, both from L. lactis, were monitored by mass spectrometry. Both proteins incorporated SeMet with high efficiencies (>90%), which greatly extends the usefulness of the expression host L. lactis for X-ray crystallography purposes. The crystal structure of ligand-free OppA was determined at 2.4 angstrom resolution by a semiautomatic approach using selenium single-wavelength anomalous diffraction phasing.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available