Development and Application of Sub-Mitochondrial Targeted Ca 2 + Biosensors.

Autor: Waldeck-Weiermair M; Molecular Biology and Biochemistry, Gottfried Schatz Research Center, Medical University of Graz, Graz, Austria., Gottschalk B; Molecular Biology and Biochemistry, Gottfried Schatz Research Center, Medical University of Graz, Graz, Austria., Madreiter-Sokolowski CT; Molecular Biology and Biochemistry, Gottfried Schatz Research Center, Medical University of Graz, Graz, Austria.; Energy Metabolism Laboratory, Institute of Translational Medicine, D-HEST, Swiss Federal Institute of Technology (ETH), Zurich, Switzerland., Ramadani-Muja J; Molecular Biology and Biochemistry, Gottfried Schatz Research Center, Medical University of Graz, Graz, Austria., Ziomek G; Molecular Biology and Biochemistry, Gottfried Schatz Research Center, Medical University of Graz, Graz, Austria., Klec C; Molecular Biology and Biochemistry, Gottfried Schatz Research Center, Medical University of Graz, Graz, Austria.; Department of Internal Medicine, Division of Oncology, Medical University of Graz, Graz, Austria., Burgstaller S; Molecular Biology and Biochemistry, Gottfried Schatz Research Center, Medical University of Graz, Graz, Austria., Bischof H; Molecular Biology and Biochemistry, Gottfried Schatz Research Center, Medical University of Graz, Graz, Austria., Depaoli MR; Molecular Biology and Biochemistry, Gottfried Schatz Research Center, Medical University of Graz, Graz, Austria., Eroglu E; Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, United States., Malli R; Molecular Biology and Biochemistry, Gottfried Schatz Research Center, Medical University of Graz, Graz, Austria.; BioTechMed-Graz, Graz, Austria., Graier WF; Molecular Biology and Biochemistry, Gottfried Schatz Research Center, Medical University of Graz, Graz, Austria.; BioTechMed-Graz, Graz, Austria.
Jazyk: angličtina
Zdroj: Frontiers in cellular neuroscience [Front Cell Neurosci] 2019 Oct 04; Vol. 13, pp. 449. Date of Electronic Publication: 2019 Oct 04 (Print Publication: 2019).
DOI: 10.3389/fncel.2019.00449
Abstrakt: Mitochondrial Ca 2+ uptake into the mitochondrial matrix is a well-established mechanism. However, the sub-organellar Ca 2+ kinetics remain elusive. In the present work we identified novel site-specific targeting sequences for the intermembrane space (IMS) and the cristae lumen (CL). We used these novel targeting peptides to develop green- and red- Ca 2+ biosensors targeted to the IMS and to the CL. Based on their distinctive spectral properties, and comparable sensitivities these novel constructs were suitable to visualize Ca 2+ -levels in various (sub) compartments in a multi-chromatic manner. Functional studies that applied these new biosensors revealed that knockdown of MCU and EMRE yielded elevated Ca 2+ levels inside the CL but not the IMS in response to IP 3 -generating agonists. Knockdown of VDAC1, however, strongly impeded the transfer of Ca 2+ through the OMM while the cytosolic Ca 2+ signal remained unchanged. The novel sub-mitochondrially targeted Ca 2+ biosensors proved to be suitable for Ca 2+ imaging with high spatial and temporal resolution in a multi-chromatic manner allowing simultaneous measurements. These informative biosensors will facilitate efforts to dissect the complex sub-mitochondrial Ca 2+ signaling under (patho)physiological conditions.
(Copyright © 2019 Waldeck-Weiermair, Gottschalk, Madreiter-Sokolowski, Ramadani-Muja, Ziomek, Klec, Burgstaller, Bischof, Depaoli, Eroglu, Malli and Graier.)
Databáze: MEDLINE
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