EstG is a novel esterase required for cell envelope integrity in Caulobacter

Curr Biol. 2023 Jan 23;33(2):228-240.e7. doi: 10.1016/j.cub.2022.11.037. Epub 2022 Dec 13.

Abstract

Proper regulation of the bacterial cell envelope is critical for cell survival. Identification and characterization of enzymes that maintain cell envelope homeostasis is crucial, as they can be targets for effective antibiotics. In this study, we have identified a novel enzyme, called EstG, whose activity protects cells from a variety of lethal assaults in the ⍺-proteobacterium Caulobacter crescentus. Despite homology to transpeptidase family cell wall enzymes and an ability to protect against cell-wall-targeting antibiotics, EstG does not demonstrate biochemical activity toward cell wall substrates. Instead, EstG is genetically connected to the periplasmic enzymes OpgH and BglX, responsible for synthesis and hydrolysis of osmoregulated periplasmic glucans (OPGs), respectively. The crystal structure of EstG revealed similarities to esterases and transesterases, and we demonstrated esterase activity of EstG in vitro. Using biochemical fractionation, we identified a cyclic hexamer of glucose as a likely substrate of EstG. This molecule is the first OPG described in Caulobacter and establishes a novel class of OPGs, the regulation and modification of which are important for stress survival and adaptation to fluctuating environments. Our data indicate that EstG, BglX, and OpgH comprise a previously unknown OPG pathway in Caulobacter. Ultimately, we propose that EstG is a novel enzyme that instead of acting on the cell wall, acts on cyclic OPGs to provide resistance to a variety of cellular stresses.

Keywords: Caulobacter crescentus; Gram-negative; antibiotics; cell envelope; envelope stress; esterase; osmoregulated periplasmic glucans; peptidoglycan; transesterase.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Caulobacter crescentus* / metabolism
  • Caulobacter* / metabolism
  • Cell Membrane / metabolism
  • Cell Wall / metabolism
  • Esterases

Substances

  • Esterases
  • Anti-Bacterial Agents
  • Bacterial Proteins