Working Paper Article Version 1 This version is not peer-reviewed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.

Subject Areas

Streptococcus pneumoniae; protein purification; protein labelling; seleno-methionine; DNA-protein interactions

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