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. Author manuscript; available in PMC: 2014 Aug 16.
Published in final edited form as: J Am Chem Soc. 2013 Feb 14;135(8):2999–3010. doi: 10.1021/ja306361q

Figure 4. Nucleotides bind to the E-to-Q mutants faster than to WT SecA.

Figure 4

WT or E210Q EcSecA were added to a cuvette containing 60 nM MANT-labeled nucleotide in KEMT buffer (pH 7.6) at 25 °C, and binding was monitored using fluorescence anisotropy spectroscopy at 352 nm. The on-rate (kon) was calculated from the observed single-exponential binding curves as explained in the SI. The labels ADP and ATP refer to Mg-MANT-ADP and Mg-MANT-ATP, respectively. The inset shows a magnified view of the data for the WT enzyme.