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Estimation of SecA-attached signal peptide-binding affinity based on fluorescence anisotropy. One micromolar IANBD-labeled SP22 in TKE buffer was mixed with the indicated final concentration of SecA or chimera protein and incubated at 20 °C for 45 min, when it was subjected to fluorescence anisotropy analysis as described in Experimental Procedures. Data were fit assuming a 1:1 binding interaction.
