Backbone resonance assignments of an artificially engineered TEM-1/PSE-4 Class A β-lactamase chimera |
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Authors: | Sébastien Morin Christopher M Clouthier Sophie Gobeil Joelle N Pelletier Stéphane M Gagné |
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Institution: | 1. PROTEO, the Québec Network for Research on Protein Structure, Function and Engineering, Université Laval, Québec, Canada 2. Département de biochimie et de microbiologie, Université Laval, Québec, Canada 3. Département de chimie, Université de Montréal, Montréal, Canada 4. Département de biochimie, Université de Montréal, Montréal, Canada
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Abstract: | The rapid evolution of Class A β-lactamases, which procure resistance to an increasingly broad panel of β-lactam antibiotics, underscores the urgency to better understand the relation between their sequence variation and their structural and functional features. To date, more than 300 clinically-relevant β-lactamase variants have been reported, and this number continues to increase. With the aim of obtaining insights into the evolutionary potential of β-lactamases, an artificially engineered, catalytically active chimera of the Class A TEM-1 and PSE-4 β-lactamases is under study by kinetics and NMR. Here we report the 1H, 13C and 15N backbone resonance assignments for the 30 kDa chimera cTEM-17m. Despite its high molecular weight, the data provide evidence that this artificially-evolved chimeric enzyme is well folded. The hydrolytic activity of cTEM-17m was determined using the chromogenic substrate CENTA, with K M = 160 ± 35 μM and k cat = 20 ± 4 s?1, which is in the same range as the values for TEM-1 and PSE-4 β-lactamases. |
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