Surface sites for engineering allosteric control in proteins

Science. 2008 Oct 17;322(5900):438-42. doi: 10.1126/science.1159052.

Abstract

Statistical analyses of protein families reveal networks of coevolving amino acids that functionally link distantly positioned functional surfaces. Such linkages suggest a concept for engineering allosteric control into proteins: The intramolecular networks of two proteins could be joined across their surface sites such that the activity of one protein might control the activity of the other. We tested this idea by creating PAS-DHFR, a designed chimeric protein that connects a light-sensing signaling domain from a plant member of the Per/Arnt/Sim (PAS) family of proteins with Escherichia coli dihydrofolate reductase (DHFR). With no optimization, PAS-DHFR exhibited light-dependent catalytic activity that depended on the site of connection and on known signaling mechanisms in both proteins. PAS-DHFR serves as a proof of concept for engineering regulatory activities into proteins through interface design at conserved allosteric sites.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allosteric Regulation
  • Allosteric Site
  • Binding Sites
  • Catalysis
  • Cryptochromes
  • Escherichia coli / enzymology
  • Flavoproteins / chemistry*
  • Flavoproteins / metabolism
  • Kinetics
  • Ligands
  • Light
  • Models, Molecular
  • NADP / metabolism
  • Protein Conformation
  • Protein Engineering*
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Recombinant Fusion Proteins / chemistry*
  • Recombinant Fusion Proteins / metabolism*
  • Tetrahydrofolate Dehydrogenase / chemistry*
  • Tetrahydrofolate Dehydrogenase / metabolism

Substances

  • Cryptochromes
  • Flavoproteins
  • Ligands
  • Recombinant Fusion Proteins
  • NADP
  • Tetrahydrofolate Dehydrogenase