ATP sequestration by a synthetic ATP-binding protein leads to novel phenotypic changes in Escherichia coli

Shaleen B. Korch, Joshua M. Stomel, Megan A. León, Matt A. Hamada, Christine R. Stevenson, Brent W. Simpson, Sunil K. Gujulla, John C. Chaput

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

Artificial proteins that bind key metabolites with high affinity and specificity hold great promise as new tools in synthetic biology, but little has been done to create such molecules and examine their effects on living cells. Experiments of this kind have the potential to expand our understanding of cellular systems, as certain phenotypes may be physically realistic but not yet observed in nature. Here, we examine the physiology and morphology of a population of Escherichia coli as they respond to a genetically encoded, non-biological ATP-binding protein. Unlike natural ATP-dependent proteins, which transiently bind ATP during metabolic transformations, the synthetic protein DX depletes the concentration of intracellular ATP and ADP by a mechanism of protein-mediated ligand sequestration. The resulting ATP/ADP imbalance leads to an adaptive response in which a large population of bacilli cells transition to a filamentous state with dense lipid structures that segregate the cells into compartmentalized units. A wide range of biochemical and microscopy techniques extensively characterized these novel lipid structures, which we have termed endoliposomes. We show that endoliposomes adopt well-defined box-like structures that span the full width of the cell but exclude the synthetic protein DX. We further show that prolonged DX exposure causes a large fraction of the population to enter a viable-but-non-culturable state that is not easily reversed. Both phenotypes correlate with strong intracellular changes in ATP and ADP concentration. We suggest that artificial proteins, such as DX, could be used to control and regulate specific targets in metabolic pathways.

Original languageEnglish (US)
Pages (from-to)451-463
Number of pages13
JournalACS Chemical Biology
Volume8
Issue number2
DOIs
StatePublished - Feb 15 2013

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

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    Korch, S. B., Stomel, J. M., León, M. A., Hamada, M. A., Stevenson, C. R., Simpson, B. W., Gujulla, S. K., & Chaput, J. C. (2013). ATP sequestration by a synthetic ATP-binding protein leads to novel phenotypic changes in Escherichia coli. ACS Chemical Biology, 8(2), 451-463. https://doi.org/10.1021/cb3004786