Twelve positions in a β-Lactamase that can expand its substrate spectrum with a single amino acid substitution

Hyojeong Yi, Kwang Hwi Cho, Yun Sung Cho, Karan Kim, William C. Nierman, Heenam Kim

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The continuous evolution of β-lactamases resulting in bacterial resistance to β-lactam antibiotics is a major concern in public health, and yet the underlying molecular basis or the pattern of such evolution is largely unknown. We investigated the mechanics of the substrate fspectrum expansion of the class A β-lactamase using PenA of Burkholderia thailandensis as a model. By analyzing 516 mutated enzymes that acquired the ceftazidime-hydrolyzing activity, we found twelve positions with single amino acid substitutions (altogether twenty-nine different substitutions), co-localized at the active-site pocket area. The ceftazidime MIC (minimum inhibitory concentration) levels and the relative frequency in the occurrence of substitutions did not correlate well with each other, and the latter appeared be largely influenced by the intrinsic mutational biases present in bacteria. Simulation studies suggested that all substitutions caused a congruent effect, expanding the space in a conserved structure called the omega loop, which in turn increased flexibility at the active site. A second phase of selection, in which the mutants were placed under increased antibiotic pressure, did not result in a second mutation in the coding region, but a mutation that increased gene expression arose in the promoter. This result suggests that the twelve amino acid positions and their specific substitutions in PenA may represent a comprehensive repertoire of the enzyme's adaptability to a new substrate. These mapped substitutions represent a comprehensive set of general mechanical paths to substrate spectrum expansion in class A β-lactamases that all share a functional evolutionary mechanism using common conserved residues.

Original languageEnglish
Article numbere37585
JournalPLoS One
Volume7
Issue number5
DOIs
Publication statusPublished - 2012 May 22

Fingerprint

Ceftazidime
amino acid substitution
Amino Acid Substitution
active sites
Burkholderia thailandensis
Catalytic Domain
Substitution reactions
antibiotics
lactams
Burkholderia
Anti-Bacterial Agents
mutation
Amino Acids
Lactams
Mutation
Microbial Sensitivity Tests
Substrates
Enzymes
enzymes
minimum inhibitory concentration

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Twelve positions in a β-Lactamase that can expand its substrate spectrum with a single amino acid substitution. / Yi, Hyojeong; Cho, Kwang Hwi; Cho, Yun Sung; Kim, Karan; Nierman, William C.; Kim, Heenam.

In: PLoS One, Vol. 7, No. 5, e37585, 22.05.2012.

Research output: Contribution to journalArticle

Yi, Hyojeong ; Cho, Kwang Hwi ; Cho, Yun Sung ; Kim, Karan ; Nierman, William C. ; Kim, Heenam. / Twelve positions in a β-Lactamase that can expand its substrate spectrum with a single amino acid substitution. In: PLoS One. 2012 ; Vol. 7, No. 5.
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