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. Author manuscript; available in PMC: 2011 Jul 9.
Published in final edited form as: Mol Cell. 2010 Jul 9;39(1):48–58. doi: 10.1016/j.molcel.2010.06.013

Figure 5.

Figure 5

MuB-DNA binding kinetics. (A) DNA binding was monitored after mixing ECFP-MuB or wild type MuB with Alexa-514 labeled dsDNA and ATP. Green and red lines indicate ECFP-MuB and wild type MuB, respectively. (B) DNA binding of the ATP pre-activated MuB was monitored by mixing ECFP-MuB or wild type MuB, preincubated with ATP for 10 min at 25 °C, with Alexa-514 labeled dsDNA. (C) The same experiment as in A was carried out at different concentrations of ECFP-MuB as indicated. Irrespective of the protein concentration, ECFP-MuB fluorescence quenching of about 1.5% was observed in several seconds following addition of ATP independent of DNA, as more clearly seen at a low protein concentration here. The final concentrations of reaction components in A and B were: 1 μM ECFP-MuB or wild type MuB, 10 ng/μl Alexa-514 labeled dsDNA and 2 mM ATP. All data are plotted as the ratio of fluorescent intensity at each time point to that at the first time point.