Article
Version 1
This version is not peer-reviewed
Complete Labelling of Pneumococcal DNA-Binding Proteins with Seleno-L-methionine
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. Preprints 2019, 2019050236 Lorenzo-Diaz, F.; Moreno-Córdoba, I.; Espinosa, M. Complete Labelling of Pneumococcal DNA-Binding Proteins with Seleno-L-methionine. Preprints 2019, 2019050236
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.
Comments (0)
We encourage comments and feedback from a broad range of readers. See criteria for comments and our Diversity statement.
Leave a public commentSend a private comment to the author(s)
* All users must log in before leaving a comment