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The interrelationship between α-synuclein (α-syn) and mitochondria is not clearly understood. Owing to the lack of the signal peptide and its predominant localization in the cytosol, α-syn is generally considered to affect mitochondrial function through some secondary effects. Contrary to this assumption, here, we show that a portion of α-syn is present in the membrane of mitochondria in normal dopaminergic neurons. The same profile is also found in other α-syn-positive neurons. Thus, binding to the membrane of mitochondria is the physiological nature of α-syn and might also contribute to the pathological role of this protein in the mitochondrial dysfunction in Parkinson's disease.