Tracking Proteins in the Cell’s Powerhouse

A labeling and electroporation workflow brings high-resolution NMR into intact organelles

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Researchers developed an NMR spectroscopy method to study protein structure, interactions, and stability inside intact mitochondria.
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The team delivered isotope-labeled proteins into isolated rat liver and pig brain mitochondria by electroporation, which preserved membrane potential and enzyme activity.
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Alpha-synuclein remained largely disordered in mitochondria, and its N-terminal region interacted with mitochondrial membranes.
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The A30P variant and methionine oxidation each reduced alpha-synuclein’s interaction with mitochondrial membranes compared with unmodified protein.
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GB1 was more stable and a GB1 variant bound more strongly inside mitochondria than in buffer, although experiments used isolated mitochondria below physiological temperatures.

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