Backbone resonance assignments of an artificially engineered TEM-1/PSE-4 Class A β-lactamase chimera.
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 (1)H, (13)C and (15)N 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.
Morin
Sébastien
S
PROTEO, the Québec Network for Research on Protein Structure, Function and Engineering, Université Laval, Québec, Canada. sebastien.morin.1@ulaval.ca
Clouthier
Christopher M
CM
Gobeil
Sophie
S
Pelletier
Joelle N
JN
Gagné
Stéphane M
SM
D019906:Nuclear Magnetic Resonance, Biomolecular; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000737:chemistry*; D001618:beta-Lactamases / Q000737:chemistry*
D011994:Recombinant Proteins; C410213:beta-lactamase PSE-4; D001618:beta-Lactamases