Asp179 in the class A β-lactamase from Mycobacterium tuberculosis is a conserved yet not essential residue due to epistasis

Conserved residues are often considered essential for function, and substitutions in such residues are expected to have a negative influence on the properties of a protein. However, mutations in a few highly conserved residues of the beta-lactamase from Mycobacterium tuberculosis, BlaC, were shown to have no or only limited negative effect on the enzyme. One such mutant, D179N, even conveyed increased ceftazidime resistance upon bacterial cells, while displaying good activity against penicillins. The crystal structures of BlaC D179N in resting state and in complex with sulbactam reveal subtle structural changes in the Omega-loop as compared to the structure of wild-type BlaC. Introducing this mutation in four other beta-lactamases, CTX-M-14, KPC-2, NMC-A and TEM-1, resulted in decreased antibiotic resistance for penicillins and meropenem. The results demonstrate that the Asp in position 179 is generally essential for class A beta-lactamases but not for BlaC, which can be explained by the importance of the interaction with the side chain of Arg164 that is absent in BlaC. It is concluded that Asp179 though conserved is not essential in BlaC, as a consequence of epistasis.

Automated summary

Conserved residues are usually essential for protein function, but mutations in highly conserved residues of BlaC did not significantly affect the enzyme. One mutation, D179N, even increased ceftazidime resistance while maintaining good activity against penicillins. Crystal structures of BlaC D179N revealed minor structural changes compared to wild-type BlaC, and introducing this mutation in other beta-lactamases resulted in decreased antibiotic resistance. The importance of the interaction with Arg164 explains why Asp179 is not essential in BlaC.