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Abstract
To detect cellular activities deep within the body using magnetic resonance platforms, magnetosomes are the ideal model of genetically-encoded nanoparticles. These membrane-bound iron biominerals produced by magnetotactic bacteria are highly regulated by approximately 30 genes; however, the number of magnetosome genes that are essential and/or constitute the root structure upon which biominerals form is largely undefined. To examine the possibility that key magnetosome genes may interact in a foreign environment, we expressed mamI and mamL as fluorescent fusion proteins in mammalian cells. Localization and potential protein-protein interaction(s) were investigated using confocal microscopy and fluorescence correlation spectroscopy (FCS). Enhanced green fluorescent protein (EGFP)-MamI and the red fluorescent Tomato-MamL displayed distinct intracellular localization, with net-like and punctate fluorescence, respectively. Remarkably, co-expression revealed co-localization of both fluorescent fusion proteins in the same punctate pattern. An interaction between MamI and MamL was confirmed by co-immunoprecipitation. In addition, changes in EGFP-MamI distribution were accompanied by acquisition of intracellular mobility which all Tomato-MamL structures displayed. Analysis of extracts from these cells by FCS was consistent with an interaction between fluorescent fusion proteins, including an increase in particle radius. Co-localization and interaction of MamI and MamL demonstrate that select magnetosome proteins may associate in mammalian cells.
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1 Lawson Research Institute, Imaging, London, Canada (GRID:grid.415847.b) (ISNI:0000 0001 0556 2414); Western University, Medical Biophysics, London, Canada (GRID:grid.39381.30) (ISNI:0000 0004 1936 8884); Western University, Collaborative Graduate Program in Molecular Imaging, London, Canada (GRID:grid.39381.30) (ISNI:0000 0004 1936 8884)
2 McMaster University, Physics & Astronomy, Hamilton, Canada (GRID:grid.25073.33) (ISNI:0000 0004 1936 8227)
3 Lawson Research Institute, Imaging, London, Canada (GRID:grid.415847.b) (ISNI:0000 0001 0556 2414)
4 McMaster University, Physics & Astronomy, Hamilton, Canada (GRID:grid.25073.33) (ISNI:0000 0004 1936 8227); McMaster University, Biochemistry & Biomedical Sciences, Hamilton, Canada (GRID:grid.25073.33) (ISNI:0000 0004 1936 8227)
5 Lawson Research Institute, Imaging, London, Canada (GRID:grid.415847.b) (ISNI:0000 0001 0556 2414); Western University, Medical Biophysics, London, Canada (GRID:grid.39381.30) (ISNI:0000 0004 1936 8884); Western University, Medical Imaging, London, Canada (GRID:grid.39381.30) (ISNI:0000 0004 1936 8884); Western University, Physics & Astronomy, London, Canada (GRID:grid.39381.30) (ISNI:0000 0004 1936 8884)
6 Lawson Research Institute, Imaging, London, Canada (GRID:grid.415847.b) (ISNI:0000 0001 0556 2414); Western University, Medical Biophysics, London, Canada (GRID:grid.39381.30) (ISNI:0000 0004 1936 8884); Western University, Collaborative Graduate Program in Molecular Imaging, London, Canada (GRID:grid.39381.30) (ISNI:0000 0004 1936 8884); Western University, Medical Imaging, London, Canada (GRID:grid.39381.30) (ISNI:0000 0004 1936 8884)
7 Lawson Research Institute, Imaging, London, Canada (GRID:grid.415847.b) (ISNI:0000 0001 0556 2414); Western University, Medical Biophysics, London, Canada (GRID:grid.39381.30) (ISNI:0000 0004 1936 8884); Western University, Collaborative Graduate Program in Molecular Imaging, London, Canada (GRID:grid.39381.30) (ISNI:0000 0004 1936 8884); Lawson Research Institute, Imaging Program, London, Canada (GRID:grid.415847.b) (ISNI:0000 0001 0556 2414)