Cell-Free Synthesis of a Functional Membrane Transporter into a Tethered Bilayer Lipid Membrane

Langmuir. 2016 Mar 15;32(10):2445-9. doi: 10.1021/acs.langmuir.5b04059. Epub 2016 Mar 2.

Abstract

Eukaryotic cell-free synthesis was used to incorporate the large and complex multispan plant membrane transporter Bot1 in a functional form into a tethered bilayer lipid membrane. The electrical properties of the protein-functionalized tethered bilayer were measured using electrochemical impedance spectroscopy and revealed a pH-dependent transport of borate ions through the protein. The efficacy of the protein synthesis has been evaluated using immunoblot analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Boric Acids / metabolism
  • Cell-Free System / chemistry
  • Cell-Free System / metabolism*
  • Dielectric Spectroscopy
  • Esters
  • Hordeum
  • Hydrogen-Ion Concentration
  • Immunoblotting
  • Lipid Bilayers / chemistry*
  • Membrane Transport Proteins / biosynthesis*
  • Membrane Transport Proteins / chemistry
  • Phosphatidylcholines / chemistry
  • Plant Proteins / biosynthesis*
  • Plant Proteins / chemistry
  • RNA, Messenger / genetics
  • RNA, Plant / genetics
  • Thioctic Acid / chemistry

Substances

  • Boric Acids
  • Esters
  • Lipid Bilayers
  • Membrane Transport Proteins
  • Phosphatidylcholines
  • Plant Proteins
  • RNA, Messenger
  • RNA, Plant
  • 1,2-diphytanoylphosphatidylcholine
  • Thioctic Acid
  • boric acid