Binding of the immunomodulatory drug Bz-423 to mitochondrial F0F1-ATP synthase in living cells by FRET acceptor photobleaching

Ilka Starke, Kathryn M. Johnson, Jan Petersen, Peter Gräber, Anthony W. Opipari, Gary D. Glick, Michael Börsch

Research output: Chapter in Book/Report/Conference proceedingConference PaperResearch

6 Citations (Scopus)

Abstract

Bz-423 is a promising new drug for treatment of autoimmune diseases. This small molecule binds to subunit OSCP of the mitochondrial enzyme FoF1-ATP synthase and modulates its catalytic activities. We investigate the binding of Bz-423 to mitochondria in living cells and how subunit rotation in FoF1-ATP synthase, i.e. the mechanochemical mechanism of this enzyme, is affected by Bz-423. Therefore, the enzyme was marked selectively by genetic fusion with the fluorescent protein EGFP to the C terminus of subunit γ. Imaging the threedimensional arrangement of mitochondria in living yeast cells was possible at superresolution using structured illumination microscopy, SIM. We measured uptake and binding of a Cy5-labeled Bz-423 derivative to mitochondrial FoF1-ATP synthase in living yeast cells using FRET acceptor photobleaching microscopy. Our data confirmed the binding of Cy5-labeled Bz-423 to the top of the F1 domain of the enzyme in mitochondria of living Saccharomyces cerevisiae cells.

Original languageEnglish
Title of host publicationMultiphoton Microscopy in the Biomedical Sciences XVI
PublisherSPIE
Number of pages11
Volume9712
ISBN (Electronic)9781628419467
DOIs
Publication statusPublished - 2016
EventMultiphoton Microscopy in the Biomedical Sciences XVI - The Moscone Center, San Francisco, United States of America
Duration: 14 Feb 201616 Feb 2016
Conference number: 17th
https://spie.org/PW16B/conferencedetails/multiphoton-microscopy

Conference

ConferenceMultiphoton Microscopy in the Biomedical Sciences XVI
Abbreviated titleSPIE BIOS 2016
CountryUnited States of America
CitySan Francisco
Period14/02/1616/02/16
Internet address

Keywords

  • acceptor photobleaching
  • Bz-423
  • FF-ATP synthase
  • FRET
  • live cell imaging
  • structured illumination microscopy

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