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In Vitro Reconstitution of Self-Organizing Protein Patterns on Supported Lipid Bilayers
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Protein Translocation Activity in Surface-Supported Lipid Bilayers.

Kanokporn Chattrakun, David P Hoogerheide1, Chunfeng Mao

  • 1Center for Neutron Research , National Institute of Standards and Technology , Gaithersburg , Maryland 20899 , United States.

Langmuir : the ACS Journal of Surfaces and Colloids
|August 27, 2019
PubMed
Summary
This summary is machine-generated.

Surface-supported lipid bilayers maintain polypeptide translocation activity, crucial for membrane protein studies. Activity depends on surface topography, impacting biochemical measurements in nanoscience applications.

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Area of Science:

  • Biophysics
  • Nanoscience
  • Biochemistry

Background:

  • Surface-supported lipid bilayers mimic cellular membranes for nanoscience studies.
  • Atomic force microscopy (AFM) and other techniques question biochemical activity preservation near surfaces.

Purpose of the Study:

  • Quantify the activity of the Escherichia coli Sec translocase within surface-supported lipid bilayers.
  • Assess how surface properties influence translocase biochemical activity and polypeptide translocation.

Main Methods:

  • Utilized two biochemical assays: ATP hydrolysis and protease protection for polypeptide translocation.
  • Employed neutron reflectometry and AFM for structural characterization.
  • Compared activity on different surfaces like glass and mica.

Main Results:

  • Sec translocase exhibited activation levels similar to solution experiments, with some ATPase populations showing hysteresis.
  • Translocation turnover numbers were comparable to solution, but the rate constant was reduced tenfold.
  • Chemomechanical coupling varied only twofold on glass compared to solution.
  • Activity was influenced by surface topography, favoring rough glass over flat mica.

Conclusions:

  • Polypeptide translocation activity is maintained in surface-adsorbed Sec systems, though with a slower rate-limiting step.
  • Surface-supported lipid bilayers can be reliably used to study membrane protein dynamics.
  • Biochemical activity measurements provide metrics for assessing the local environment of surface-supported lipid bilayers.