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SecA competes with ribosomes for binding to SecYEG reconstituted into nanodiscs. Binding of RNCs to purified SecYEG in nanodiscs in the presence of increasing concentrations of SecA with AMP-PNP. The fraction of RNC-bound SecYEG complex was determined by FCS as described under “Results.” Detergent-solubilized SecYEG primarily interacted with RNC even in a large excess of SecA (white bar).
