Version 1
: Received: 17 May 2019 / Approved: 20 May 2019 / Online: 20 May 2019 (10:03:26 CEST)
How to cite:
Lorenzo-Diaz, F.; Moreno-Córdoba, I.; Espinosa, M. Complete Labelling of Pneumococcal DNA-Binding Proteins with Seleno-L-methionine. Preprints2019, 2019050236
Lorenzo-Diaz, F.; Moreno-Córdoba, I.; Espinosa, M. Complete Labelling of Pneumococcal DNA-Binding Proteins with Seleno-L-methionine. Preprints 2019, 2019050236
Lorenzo-Diaz, F.; Moreno-Córdoba, I.; Espinosa, M. Complete Labelling of Pneumococcal DNA-Binding Proteins with Seleno-L-methionine. Preprints2019, 2019050236
APA Style
Lorenzo-Diaz, F., Moreno-Córdoba, I., & Espinosa, M. (2019). Complete Labelling of Pneumococcal DNA-Binding Proteins with Seleno-L-methionine. Preprints. https://doi.org/
Chicago/Turabian Style
Lorenzo-Diaz, F., Inmaculada Moreno-Córdoba and Manuel Espinosa. 2019 "Complete Labelling of Pneumococcal DNA-Binding Proteins with Seleno-L-methionine" Preprints. https://doi.org/
Abstract
Streptococcus pneumoniae is an pathogenic and opportunistic Gram-positive bacteria that is the leading cause of community acquired respiratory diseases, varying from mild- to deathly- infections. Appearance of antibiotic resistant isolates has prompted the search for novel targets. One of the most promising approaches is the structure-based knowledge of possible targets in conjunction to rational design and docking of inhibitors of the chosen targets. A useful technique to help solving protein structures is to label them with a heavy atom, like selenium, that facilitates tracing of the some of the amino acid residues. We have chosen two pneumococcal DNA-binding proteins, namely the relaxase domain of MobM protein from plasmid pMV158, and the RelB-RelE antitoxin-toxin protein complex. Through the update of a previous protocol [1] that uses seleno-L-methionine, we could achieve 100% labelling of the proteins. Furthermore, the labelled proteins retained full activity as judged from relaxation of supercoiled plasmid DNA and from gel-retardation assays.
Keywords
Streptococcus pneumoniae; protein purification; protein labelling; seleno-methionine; DNA-protein interactions
Subject
Biology and Life Sciences, Biochemistry and Molecular Biology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
