Comparative studies on detergent-assisted apocytochrome b6 reconstitution into liposomal bilayers monitored by Zetasizer instruments

PLoS One. 2014 Nov 25;9(11):e111341. doi: 10.1371/journal.pone.0111341. eCollection 2014.

Abstract

The present paper is a systematic, comparative study on the reconstitution of an apocytochrome b6 purified from a heterologous system using a detergent-free method and reconstitution into liposomes performed using three different detergents: SDS, Triton X-100 and DM, and two methods of detergent removal by dialysis and using Bio-Beads. The product size, its distribution and zeta potential, and other parameters were monitored throughout the process. We found that zeta potential of proteoliposomes is correlated with reconstitution efficiency and, as such, can serve as a quick and convenient quality control for reconstitution experiments. We also advocate using detergent-free protein purification methods as they allow for an unfettered choice of detergent for reconstitution, which is the most crucial factor influencing the final product parameters.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cytochromes b6 / chemistry*
  • Detergents / chemistry
  • Lipid Bilayers / chemistry*
  • Liposomes / chemistry*
  • Membrane Potentials

Substances

  • Cytochromes b6
  • Detergents
  • Lipid Bilayers
  • Liposomes

Grants and funding

This research was supported by the University of Wrocław, Poland to AS. Publication fee was partially paid by the Wrocław Center for Biotechnology program KNOW (National Scientific Leadership Center) 2014-2018.