Engineered proteins with sensing and activating modules for automated reprogramming of cellular functions

Nat Commun. 2017 Sep 7;8(1):477. doi: 10.1038/s41467-017-00569-6.

Abstract

Protein-based biosensors or activators have been engineered to visualize molecular signals or manipulate cellular functions. Here we integrate these two functionalities into one protein molecule, an integrated sensing and activating protein (iSNAP). A prototype that can detect tyrosine phosphorylation and immediately activate auto-inhibited Shp2 phosphatase, Shp2-iSNAP, is designed through modular assembly. When Shp2-iSNAP is fused to the SIRPα receptor which typically transduces anti-phagocytic signals from the 'don't eat me' CD47 ligand through negative Shp1 signaling, the engineered macrophages not only allow visualization of SIRPα phosphorylation upon CD47 engagement but also rewire the CD47-SIRPα axis into the positive Shp2 signaling, which enhances phagocytosis of opsonized tumor cells. A second SIRPα Syk-iSNAP with redesigned sensor and activator modules can likewise rewire the CD47-SIRPα axis to the pro-phagocytic Syk kinase activation. Thus, our approach can be extended to execute a broad range of sensing and automated reprogramming actions for directed therapeutics.Protein-based biosensors have been engineered to interrogate cellular signaling and manipulate function. Here the authors demonstrate iSNAP, a tool to detect tyrosine phosphorylation and activate desired protein enzymes allowing the control of phagocytosis in macrophages.

Publication types

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

MeSH terms

  • Animals
  • Antigens, Differentiation / genetics
  • Antigens, Differentiation / metabolism
  • CD47 Antigen / genetics
  • CD47 Antigen / metabolism*
  • Cells, Cultured
  • Female
  • Fibroblasts / metabolism
  • Fluorescence Resonance Energy Transfer
  • Humans
  • Macrophages / metabolism*
  • Mice, Inbred C57BL
  • Phagocytosis
  • Phosphorylation
  • Protein Engineering / methods*
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11 / genetics
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11 / metabolism
  • Receptors, Immunologic / genetics
  • Receptors, Immunologic / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism*
  • Signal Transduction
  • Syk Kinase / genetics
  • Syk Kinase / metabolism
  • Tyrosine / metabolism*

Substances

  • Antigens, Differentiation
  • CD47 Antigen
  • Receptors, Immunologic
  • Recombinant Proteins
  • SIRPA protein, human
  • Tyrosine
  • Syk Kinase
  • Syk protein, mouse
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11