Partial Complex I Inhibition Decreases Mitochondrial Motility and Increases Matrix Protein Diffusion As Revealed by Fluorescence Correlation Spectroscopy.
From: Department of Membrane Biochemistry, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands.
Biochimica et biophysica acta
- Publish Date: Jul 2007
- ISSN: 0006-3002
- Volume: 1767
- Issue: 7
- Pages: 940-7
- Medium: Print
- Language: English
- Citation (JAMA): Koopman Werner J H, Hink Mark A, Verkaart Sjoerd, et al. Partial Complex I Inhibition Decreases Mitochondrial Motility and Increases Matrix Protein Diffusion As Revealed by Fluorescence Correlation Spectroscopy.. Biochim. Biophys. Acta Jul 2007;1767:940-7
Abstract
We previously reported that inhibition of mitochondrial complex I (CI) by rotenone induces marked increases in mitochondrial length and degree of branching, thus revealing a relationship between mitochondrial function and shape. We here describe the first time use of fluorescence correlation spectroscopy (FCS) to simultaneously probe mitochondrial mobility and intra-matrix protein diffusion, with the aim to investigate the effects of chronic CI inhibition on the latter two parameters. To this end, EYFP was expressed in the mitochondrial matrix of human skin fibroblasts (mitoEYFP) using baculoviral transduction and its diffusion monitored by FCS. This approach revealed the coexistence of moving and stationary mitochondria within the same cell and enabled simultaneous quantification of mitochondrial velocity and mitoEYFP diffusion. When CI activity was chronically reduced by 80% using rotenone treatment, the percentage of moving mitochondria and their velocity decreased by 30%. MitoEYFP diffusion did not differ between moving and stationary mitochondria but was increased 2-fold in both groups of mitochondria following rotenone treatment. We propose that the increase in matrix protein diffusion together with the increase in mitochondrial length and degree of branching constitutes part of an adaptive response which serves to compensate for the reduction in CI activity and mitochondrial motility.
Mesh Headings (Keywords): Bacterial Proteins, Baculoviridae, Cells, Cultured, Electron Transport Complex I, Fibroblasts, Humans, Luminescent Proteins, Microscopy, Fluorescence, Mitochondria, Mitochondrial Proteins, Rotenone, Skin, Spectrometry, Fluorescence, Uncoupling Agents
Check for Full Text / PubMed Unique Identifier (PMID): 17490603
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