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Molecular mechanism of J-Domain-Triggered ATP hydrolysis by Hsp70 chaperones

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Methods : Isolated rat brain mitochondria suspended in 50mM phosphate buffer, pH 7.4 (200l containing 200g protein) were incubated with varying concentrations of DA (100400M) for up to 2 hours with or without additions such as reduced glutathione (5mM), MitoQ (50nM), catalase (50g/ml), mannitol (20mM), DMSO (20mM) at 37C in 50 mM phosphate buffer, pH 7.4 in a total volume of 400 l