The Structural Bases of Antibiotic Resistance in the Clinically Derived Mutant β-Lactamases TEM-30, TEM-32, and TEM-34*

By Xiaojun Wang, G. Minasov, Brian K. Shoichet. 2002

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Widespread use of β-lactam antibiotics has promoted the evolution of β-lactamase mutant enzymes that can hydrolyze ever newer classes of these drugs. Among the most pernicious mutants are the inhibitor-resistant TEM β-lactamases (IRTs), which elude mechanism-based inhibitors, such as clavulanate. Despite much research on these IRTs, little is known about the structural bases of their action. This has made it difficult to understand how many of the resistance substitutions act as they often occur far from Ser-130. Here, three IRT structures, TEM-30 (R244S), TEM-32 (M69I/M182T), and TEM-34 (M69V), are determined by x-ray crystallography at 2.00, 1.61, and 1.52 Å, respectively. In TEM-30, the Arg-244 → Ser substitution (7.8 Å from Ser-130) displaces a conserved water molecule that usually interacts with the β-lactam C3 carboxylate. In TEM-32, the substitution Met-69 → Ile (10 Å from Ser-130) appears to distort Ser-70, which in turn causes Ser-130 to adopt a new conformation, moving its Oγ further away, 2.3 Å from where the inhibitor would bind. This substitution also destabilizes the enzyme by 1.3 kcal/mol. The Met-182 → Thr substitution (20 Å from Ser-130) has no effect on enzyme activity but rather restabilizes the enzyme by 2.9 kcal/mol. In TEM-34, the Met-69 → Val substitution similarly leads to a conformational change in Ser-130, this time causing it to hydrogen bond with Lys-73 and Lys-234. This masks the lone pair electrons of Ser-130 Oγ, reducing its nucleophilicity for cross-linking. In these three structures, distant substitutions result in accommodations that converge on the same point of action, the local environment of Ser-130.

Subfields: Computational Theory and Mathematics, Molecular Medicine, Pharmacology

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Citations in the corpus

Three Decades of β-Lactamase Inhibitors (2010, background): “, compensates for the loss of the Ser130 hydroxyl group by donating a proton to the -lactam nitrogen (153, 157, 180, 408, 438).”