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FEBS Lett. 2003 Nov 6;554(1-2):194-200.

Photoconversion of matrix targeted GFP enables analysis of continuity and intermixing of the mitochondrial lumen.

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  • 1Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany. sjakobs@gwdg.de


We establish photoconversion of green fluorescent protein (GFP) as an optical 'highlighter' to investigate the continuity of the mitochondrial matrix in living budding yeast (Saccharomyces cerevisiae). Photoconversion of GFP resulting in a marked shift of the absorption and emission spectra to longer wavelengths is elicited, under low oxygen conditions, by irradiation with blue light. Photoconversion induced a several 100-fold increase in red fluorescence of matrix targeted GFP without affecting cell viability. The color changing facilitates simple and effective regional optical marking in a conventional fluorescence microscope. We found the mitochondrial compartment of S. cerevisiae to generally consist of one luminally continuous large part and occasionally some additional smaller fragments. Separated fragments fuse within a few minutes to the large part, resulting in a rapid intermixing of the entire mitochondrial matrix compartment. In Deltafis1 and Deltadnm1 mutants restricted in outer membrane fission, the mitochondria are still luminally continuous, suggesting a tight coupling of inner and outer membrane fissions. Matrix constrictions frequently occurring in wild type cells as well as in Deltafis1 and Deltadnm1 mutants do not interfere with luminal continuity.

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