title: 15N, 13C and 1H backbone resonance assignments of an artificially engineered TEM-1/PSE-4 class A β-lactamase chimera and its deconvoluted mutant.
abstract: 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.
authors.affiliation: Department of Biochemistry, Université de Montréal, Montréal, QC, Canada.|PROTEO, The Québec Network for Research on Protein Function, Engineering and Applications, Québec, QC, Canada.
authors.forename: Sophie M C
authors.initials: SM
authors.lastname: Gobeil
authors.affiliation: PROTEO, The Québec Network for Research on Protein Function, Engineering and Applications, Québec, QC, Canada.|INRS-Institut Armand-Frappier, Université du Québec, Québec, QC, Canada.|GRASP, Groupe de Recherche Axé sur la Structure des Protéines, McGill University, Montréal, QC, Canada.|Structural Biology Initiative, CUNY Advanced Science Research Center, New York, NY, USA.
authors.forename: Donald
authors.initials: D
authors.lastname: Gagné
authors.affiliation: PROTEO, The Québec Network for Research on Protein Function, Engineering and Applications, Québec, QC, Canada.|INRS-Institut Armand-Frappier, Université du Québec, Québec, QC, Canada.|GRASP, Groupe de Recherche Axé sur la Structure des Protéines, McGill University, Montréal, QC, Canada.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Doucet
authors.affiliation: Department of Biochemistry, Université de Montréal, Montréal, QC, Canada. joelle.pelletier@umontreal.ca.|PROTEO, The Québec Network for Research on Protein Function, Engineering and Applications, Québec, QC, Canada. joelle.pelletier@umontreal.ca.|Department of Chemistry, Université de Montréal, Montréal, Canada. joelle.pelletier@umontreal.ca.
authors.forename: Joelle N
authors.initials: JN
authors.lastname: Pelletier
mesh_terms: D002247:Carbon Isotopes; D050505:Mutant Proteins / Q000737:chemistry*; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular; D015202:Protein Engineering; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry*; D014316:Tritium; D001618:beta-Lactamases / Q000737:chemistry*
chemical_list: D002247:Carbon Isotopes; D050505:Mutant Proteins; D009587:Nitrogen Isotopes; D011994:Recombinant Proteins; D014316:Tritium; C064821:beta-lactamase PSE-1; D001618:beta-Lactamases; C034413:beta-lactamase TEM-1
keywords: Antibiotic resistance; Chimera; PSE-4; Protein engineering; TEM-1; β-Lactamase