Leveraging Lipid-Protein Interactions to Engineer Spatial Organization in Cell-free Systems

NU 2021-231

INVENTORS
  • Neha Kamat*
  • Justin Alexander Peruzzi
  • Jan Steinkuehler

SHORT DESCRIPTION

This innovative method enables the control of membrane protein location and distribution in synthetic lipid membranes using lipid-protein interactions, without chemical conjugation, facilitating the development of advanced cell-free systems.

BACKGROUND

Spatially controlled organization of membrane proteins in synthetic systems poses significant challenges, with current technologies relying heavily on chemical conjugations or fusions with biological components. Existing methods lack the ability to spontaneously integrate proteins into synthetic vesicle membranes, limiting the potential for creating complex, functional cell-free systems.

ABSTRACT

This technology leverages the interplay between lipid membranes and proteins to control protein distribution in synthetic lipid vesicles. Utilizing cell-free protein expression systems, proteins are synthesized and directed into membrane environments that are hydrophobically matched, enhancing protein integration. This approach bypasses traditional chemical conjugation methods, employing lipid vesicles with varying hydrophobic thicknesses to direct protein localization. By utilizing computational modeling and experimental design, this method allows for the one-pot synthesis of proteins, creating vesicles with controlled protein distributions. The workflow involves de novo protein design and molecular dynamics simulation, offering a systematic approach to study membrane protein biophysics, potentially revolutionizing the design of membrane-based materials.

APPLICATIONS

  • Self-assembly in cell-free systems
  • Dynamic protein scaffolds
  • Vesicle-based Biosensor
ADVANTAGES
  • Spontaneous integration
  • One-pot synthesis

PUBLICATIONS

Neha P. Kamat et al, "Hydrophobic mismatch drives self-organization of designer proteins into synthetic membranes" Nature Communications, 11 April 2024.

IP STATUS

US Patent Pending

Patent Information: