Supplementary Materials

Ras activation by SOS: Allosteric regulation by altered fluctuation dynamics

Lars Iversen, Hsiung-Lin Tu, Wan-Chen Lin, Sune M. Christensen, Steven M. Abel, Jeff Iwig, Hung-Jen Wu, Jodi Gureasko, Christopher Rhodes, Rebecca S. Petit, Scott D. Hansen, Peter Thill, Cheng-Han Yu, Dimitrios Stamou, Arup K. Chakraborty, John Kuriyan, Jay T. Groves

Materials/Methods, Supplementary Text, Tables, Figures, and/or References

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  • Materials and Methods
  • Figs. S1 to S18
  • Tables S1 to S4
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Images, Video, and Other Other Media

Movie S1
Epifluorescence time lapse of SOScat-catalyzed nucleotide exchange reactions in the corral assay. Immediately prior to capturing the movie, SOScat was injected as a pulse through the flow chamber. Flowing in non-fluorescent GTP then initiated the reaction, and the time-lapse imaging was started. Individual corrals are observed to gradually turn dark, as SOS within the corral exchanges fluorescent nucleotide with non-fluorescent nucleotide. The difference in fluorescence decay kinetics between corrals is visible by eye. The time interval per frame is 35 s (0.0286 Hz) and the total duration of the movie is 1365 s. The movie was recorded with 400 msec exposure time. Individual frames were aligned using the StackReg command of ImageJ.
Movie S2
TIRFM single molecule tracking of SOScat-Atto647N in corrals after the reaction. False color overlay showing the epifluorescence emission from Ras-GTP-488 in red and the TIRFM emission from SOScat-Atto647N in green. A clear correspondence between dark corrals and SOScat enzymes is observed. The movie was recorded at ~5.5 Hz with 150 msec exposure time. The duration of movie is ~3.6 s (20 frames).