15N, 13C and 1H backbone resonance assignments of an artificially engineered TEM-1/PSE-4 class A β-lactamase chimera and its deconvoluted mutant.

Sophie M C Gobeil (1, 2, 3) , Donald Gagné (4, 5, 3) , Nicolas Doucet (2, 4, 6) , Joelle N Pelletier (1, 3)

1 UdeM - Université de Montréal
2 ULaval - Université Laval [Québec]
3 The Québec Network for Research on Protein Function, Engineering and Applications
4 INRS-IAF - Institut Armand Frappier
5 The Graduate Center - CUNY Graduate Center
6 McGill University = Université McGill [Montréal, Canada]

The widespread use of β-lactam antibiotics has given rise to a dramatic increase in clinically-relevant β-lactamases. Understanding the structure/function relation in these variants is essential to better address the ever-growing incidence of antibiotic resistance. We previously reported the backbone resonance assignments of a chimeric protein constituted of segments of the class A β-lactamases TEM-1 and PSE-4 (Morin et al. in Biomol NMR Assign 4:127-130, 2010. doi: 10.1007/s12104-010-9227-8 ). That chimera, cTEM17m, held 17 amino acid substitutions relative to TEM-1 β-lactamase, resulting in a well-folded and fully functional protein with increased dynamics. Here we report the (1)H, (13)C and (15)N backbone resonance assignments of chimera cTEM-19m, which includes 19 substitutions and exhibits increased active-site perturbation, as well as one of its deconvoluted variants, as the first step in the analysis of their dynamic behaviours.

Mots clés: Antibiotic resistance; Chimera; PSE-4; Protein engineering; TEM-1; β-Lactamase

Domaines: Sciences du Vivant [q-bio]

Sophie M C Gobeil, Donald Gagné, Nicolas Doucet, Joelle N Pelletier. 15N, 13C and 1H backbone resonance assignments of an artificially engineered TEM-1/PSE-4 class A β-lactamase chimera and its deconvoluted mutant.. Biomolecular NMR Assignments, 2016, 10 (1), pp.93-9. ⟨10.1007/s12104-015-9645-8⟩. ⟨pasteur-01351188⟩