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Computational design of a Zn2+ receptor that controls bacterial gene expression.

Publication ,  Journal Article
Dwyer, MA; Looger, LL; Hellinga, HW
Published in: Proc Natl Acad Sci U S A
September 30, 2003

The control of cellular physiology and gene expression in response to extracellular signals is a basic property of living systems. We have constructed a synthetic bacterial signal transduction pathway in which gene expression is controlled by extracellular Zn2+. In this system a computationally designed Zn2+-binding periplasmic receptor senses the extracellular solute and triggers a two-component signal transduction pathway via a chimeric transmembrane protein, resulting in transcriptional up-regulation of a beta-galactosidase reporter gene. The Zn2+-binding site in the designed receptor is based on a four-coordinate, tetrahedral primary coordination sphere consisting of histidines and glutamates. In addition, mutations were introduced in a secondary coordination sphere to satisfy the residual hydrogen-bonding potential of the histidines coordinated to the metal. The importance of the secondary shell interactions is demonstrated by their effect on metal affinity and selectivity, as well as protein stability. Three designed protein sequences, comprising two distinct metal-binding positions, were all shown to bind Zn2+ and to function in the cell-based assay, indicating the generality of the design methodology. These experiments demonstrate that biological systems can be manipulated with computationally designed proteins that have drastically altered ligand-binding specificities, thereby extending the repertoire of genetic control by extracellular signals.

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Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

September 30, 2003

Volume

100

Issue

20

Start / End Page

11255 / 11260

Location

United States

Related Subject Headings

  • Zinc
  • Temperature
  • Signal Transduction
  • Receptors, Cell Surface
  • Protein Denaturation
  • Mutagenesis
  • Models, Molecular
  • Gene Expression Regulation, Bacterial
 

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Dwyer, M. A., Looger, L. L., & Hellinga, H. W. (2003). Computational design of a Zn2+ receptor that controls bacterial gene expression. Proc Natl Acad Sci U S A, 100(20), 11255–11260. https://doi.org/10.1073/pnas.2032284100
Dwyer, M. A., L. L. Looger, and H. W. Hellinga. “Computational design of a Zn2+ receptor that controls bacterial gene expression.Proc Natl Acad Sci U S A 100, no. 20 (September 30, 2003): 11255–60. https://doi.org/10.1073/pnas.2032284100.
Dwyer MA, Looger LL, Hellinga HW. Computational design of a Zn2+ receptor that controls bacterial gene expression. Proc Natl Acad Sci U S A. 2003 Sep 30;100(20):11255–60.
Dwyer, M. A., et al. “Computational design of a Zn2+ receptor that controls bacterial gene expression.Proc Natl Acad Sci U S A, vol. 100, no. 20, Sept. 2003, pp. 11255–60. Pubmed, doi:10.1073/pnas.2032284100.
Dwyer MA, Looger LL, Hellinga HW. Computational design of a Zn2+ receptor that controls bacterial gene expression. Proc Natl Acad Sci U S A. 2003 Sep 30;100(20):11255–11260.
Journal cover image

Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

September 30, 2003

Volume

100

Issue

20

Start / End Page

11255 / 11260

Location

United States

Related Subject Headings

  • Zinc
  • Temperature
  • Signal Transduction
  • Receptors, Cell Surface
  • Protein Denaturation
  • Mutagenesis
  • Models, Molecular
  • Gene Expression Regulation, Bacterial