7A74

Structure of G132N BlaC from Mycobacterium tuberculosis

Summary for 7A74

• Entry DOI: 10.2210/pdb7a74/pdb
• Descriptor: Beta-lactamase, GLYCEROL, 1,2-ETHANEDIOL, ... (6 entities in total)
• Functional Keywords: blac, beta-lactamase, mycobacterium tuberculosis, hydrolase
• Biological source: Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
• Total number of polymer chains: 1
• Total formula weight: 29880.11

Authors

Chikunova, A.,Ahmad, M.U.,Perrakis, A.,Ubbink, M. (deposition date: 2020-08-27, release date: 2021-05-19, Last modification date: 2024-10-16)

Primary citation

Elings, W.,Chikunova, A.,van Zanten, D.B.,Drenth, R.,Ahmad, M.U.D.,Blok, A.J.,Timmer, M.,Perrakis, A.,Ubbink, M.
Two beta-Lactamase Variants with Reduced Clavulanic Acid Inhibition Display Different Millisecond Dynamics.
Antimicrob.Agents Chemother., 65:e0262820-e0262820, 2021

PubMed Abstract: The β-lactamase of Mycobacterium tuberculosis, BlaC, is susceptible to inhibition by clavulanic acid. The ability of this enzyme to escape inhibition through mutation was probed using error-prone PCR combined with functional screening in Escherichia coli. The variant that was found to confer the most inhibitor resistance, K234R, as well as variant G132N that was found previously were characterized using X-ray crystallography and nuclear magnetic resonance (NMR) relaxation experiments to probe structural and dynamic properties. The G132N mutant exists in solution in two almost equally populated conformations that exchange with a rate of ca. 88 s. The conformational change affects a broad region of the enzyme. The crystal structure reveals that the Asn132 side chain forces the peptide bond between Ser104 and Ile105 in a -conformation. The crystal structure suggests multiple conformations for several side chains (e.g., Ser104 and Ser130) and a short loop (positions 214 to 216). In the K234R mutant, the active-site dynamics are significantly diminished with respect to the wild-type enzyme. These results show that multiple evolutionary routes are available to increase inhibitor resistance in BlaC and that active-site dynamics on the millisecond time scale are not required for catalytic function.

PubMed: 34031049
DOI: 10.1128/AAC.02628-20

Experimental method

X-RAY DIFFRACTION (1.6 Å)