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

doi:10.1117/12.2209645

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

Biochemistry & Molecular Biology

Research output: Chapter in Book/Report/Conference proceeding › Conference Paper › Research

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 F_oF₁-ATP synthase and modulates its catalytic activities. We investigate the binding of Bz-423 to mitochondria in living cells and how subunit rotation in F_oF₁-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 F_oF₁-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 language	English
Title of host publication	Multiphoton Microscopy in the Biomedical Sciences XVI
Publisher	SPIE - International Society for Optical Engineering
Number of pages	11
Volume	9712
ISBN (Electronic)	9781628419467
DOIs	https://doi.org/10.1117/12.2209645
Publication status	Published - 2016
Event	Multiphoton Microscopy in the Biomedical Sciences XVI - The Moscone Center, San Francisco, United States of America Duration: 14 Feb 2016 → 16 Feb 2016 Conference number: 17th https://spie.org/PW16B/conferencedetails/multiphoton-microscopy

Conference

Conference	Multiphoton Microscopy in the Biomedical Sciences XVI
Abbreviated title	SPIE BIOS 2016
Country/Territory	United States of America
City	San Francisco
Period	14/02/16 → 16/02/16
Internet address	https://spie.org/PW16B/conferencedetails/multiphoton-microscopy

Keywords

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

Access to Document

10.1117/12.2209645

Cite this

Starke, I., Johnson, K. M., Petersen, J., Gräber, P., Opipari, A. W., Glick, G. D., & Börsch, M. (2016). Binding of the immunomodulatory drug Bz-423 to mitochondrial F₀F₁-ATP synthase in living cells by FRET acceptor photobleaching. In Multiphoton Microscopy in the Biomedical Sciences XVI (Vol. 9712). Article 97120P SPIE - International Society for Optical Engineering. https://doi.org/10.1117/12.2209645

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

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.",

keywords = "acceptor photobleaching, Bz-423, FF-ATP synthase, FRET, live cell imaging, structured illumination microscopy",

author = "Ilka Starke and Johnson, \{Kathryn M.\} and Jan Petersen and Peter Gr{\"a}ber and Opipari, \{Anthony W.\} and Glick, \{Gary D.\} and Michael B{\"o}rsch",

year = "2016",

doi = "10.1117/12.2209645",

language = "English",

volume = "9712",

booktitle = "Multiphoton Microscopy in the Biomedical Sciences XVI",

publisher = "SPIE - International Society for Optical Engineering",

address = "United States of America",

note = "Multiphoton Microscopy in the Biomedical Sciences XVI, SPIE BIOS 2016 ; Conference date: 14-02-2016 Through 16-02-2016",

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Starke, I, Johnson, KM, Petersen, J, Gräber, P, Opipari, AW, Glick, GD & Börsch, M 2016, Binding of the immunomodulatory drug Bz-423 to mitochondrial F₀F₁-ATP synthase in living cells by FRET acceptor photobleaching. in Multiphoton Microscopy in the Biomedical Sciences XVI. vol. 9712, 97120P, SPIE - International Society for Optical Engineering, Multiphoton Microscopy in the Biomedical Sciences XVI, San Francisco, California, United States of America, 14/02/16. https://doi.org/10.1117/12.2209645

Binding of the immunomodulatory drug Bz-423 to mitochondrial F₀F₁-ATP synthase in living cells by FRET acceptor photobleaching. / Starke, Ilka; Johnson, Kathryn M.; Petersen, Jan et al.
Multiphoton Microscopy in the Biomedical Sciences XVI. Vol. 9712 SPIE - International Society for Optical Engineering, 2016. 97120P.

Research output: Chapter in Book/Report/Conference proceeding › Conference Paper › Research

TY - GEN

T1 - Binding of the immunomodulatory drug Bz-423 to mitochondrial F0F1-ATP synthase in living cells by FRET acceptor photobleaching

AU - Starke, Ilka

AU - Johnson, Kathryn M.

AU - Petersen, Jan

AU - Gräber, Peter

AU - Opipari, Anthony W.

AU - Glick, Gary D.

AU - Börsch, Michael

N1 - Conference code: 17th

PY - 2016

Y1 - 2016

N2 - 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.

AB - 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.

KW - acceptor photobleaching

KW - Bz-423

KW - FF-ATP synthase

KW - FRET

KW - live cell imaging

KW - structured illumination microscopy

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U2 - 10.1117/12.2209645

DO - 10.1117/12.2209645

M3 - Conference Paper

AN - SCOPUS:84975063094

VL - 9712

BT - Multiphoton Microscopy in the Biomedical Sciences XVI

PB - SPIE - International Society for Optical Engineering

T2 - Multiphoton Microscopy in the Biomedical Sciences XVI

Y2 - 14 February 2016 through 16 February 2016

ER -