Figure 5 | Scientific Reports

Figure 5

From: The uncoupled ATPase activity of the ABC transporter BtuC2D2 leads to a hysteretic conformational change, conformational memory and improved activity

Figure 5

Long-term memory of BtuC2D2.

Freshly prepared BtuC2D2. (time zero) was incubated under the working conditions at 37 °C (in the absence of BtuF). At the indicated intervals samples were withdrawn, ATP was removed by microfiltration and BtuF-binding was measured by SPR. After 5 hours ATP was removed from all of the remaining volume and the protein was incubated under the idle conditions (at 4 °C, to avoid idleness-associated detrimental effects) for another 2.5 hours, during which samples were removed and BtuF-binding was measured by SPR. Subsequently, AMP-PNP was added and BtuC2D2 was moved back to 37 °C (in the ATP-bound state the protein does not deteriorate at 37 °C). At the indicated intervals samples were withdrawn, the AMP-PNP was removed by microfiltration and BtuF-binding was measured by SPR. Results are presented as percent change from the activity of the fresh, naïve protein. Error bars represent standard deviations of triplicates from two different experiments (n = 6).

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