Computational design and biochemical characterization of maize nonspecific lipid transfer protein variants for biosensor applications

Authors

  • Eun Jung Choi,

    1. Division of Biology, California Institute of Technology, Pasadena, California 91125, USA
    Search for more papers by this author
    • These authors contributed equally to this work.

  • Jessica Mao,

    1. Division of Biology, California Institute of Technology, Pasadena, California 91125, USA
    Search for more papers by this author
    • These authors contributed equally to this work.

  • Stephen L. Mayo

    Corresponding author
    1. Howard Hughes Medical Institute, Division of Biology and Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125, USA
    • Howard Hughes Medical Institute, Division of Biology and Division of Chemistry and Chemical Engineering, California Institute of Technology, 1200 E. California Blvd., Pasadena, CA 91125, USA; fax: (626) 568-0934.
    Search for more papers by this author

Abstract

Lipid transfer proteins (LTPs) are a family of proteins that bind and transfer lipids. Utilizing the maize LTP, we have successfully engineered fluorescent reagentless biosensors for the natural ligand of LTPs; this was achieved by using computational protein design to remove a disulfide bridge and attaching a thio-reactive fluorophore. Conformational change induced by ligand titration is thought to affect the fluorescence of the fluorophore, allowing detection of ligand binding. Fluorescence measurements show that our LTP variants have affinity to palmitate that is consistent with wild-type LTP. These molecules have the potential to be utilized as scaffolds to design hydrophobic ligand biosensors or to serve as drug carriers.

Ancillary