Spatial Regulators for Bacterial Cell Division Self-Organize into Surface Waves in Vitro
Martin Loose, Elisabeth Fischer-Friedrich, Jonas Ries, Karsten Kruse, Petra Schwille

Supporting Online Material

This supplement contains:
Materials and Methods
SOM Text
Figs. S1 to S13
Table S1
References

This file is in Adobe Acrobat PDF format.

Other Supporting Online Material for this manuscript includes the following: (available at www.sciencemag.org/cgi/content/full/320/5877/789/DC1)
Movies S1 to S13

Movie s1
Movie of Min protein waves. MinD (1 μM), doped with 20% Bodipy-labeled MinD (green), MinE (1 μM), doped with 10% Alexa647-labeled MinE (red). Scale bar is 50 μm, frames are 9 s apart.

Movie s2
Movie of Min protein waves, scale bar is 50 μm, frames are 13 s apart. MinD (1 μM), doped with 20% Bodipy-labeled MinD (green), MinE (1.5 μM), doped with 10% Alexa647-labeled MinE (red).

Movie s3
Example of protein dynamics in the presence of 0.125 μM MinE, doped with 10% Alexa647-labeled MinE (red) and 1 μM MinD, doped with 20% Bodipy-labeled MinD (green). Scale bar is 50 μm, frames are 18 s apart.

Movie s4
Movie of initiation of Min protein waves. MinD (1 μM), doped with 20% Bodipylabeled MinD (green), MinE (1 μM), doped with 10% Alexa647-labeled MinE (red). Scale bar is 20 μm, frames are 23 s apart.

Movie s5
Movie of synchronization of Min protein waves. MinD (1 μM), doped with 20% Bodipy-labeled MinD (green), MinE (1 μM), doped with 10% Alexa647-labeled MinE (red). Scale bar is 50 μm, frames are 13 s apart. MinD (green), MinE (red).

Movie s6
Movie of a spiral of Min proteins. MinD (1 μM), MinE (1 μM), doped with 10% Alexa647-labeled MinE. Scale bar is 100 μm, frames are 14 s apart.

Movie s7
Movie of a spiral of Min proteins. MinD (1 μM), doped with 20% Bodipy-labeled MinD (green), MinE (1 μM), doped with 10% Alexa647-labeled MinE (red). Scale bar is 50 μm, frames are 10 s apart.

Movie s8
Movie of a spiral of Min proteins. MinD (1 μM), doped with 20% Bodipy-labeled MinD (green), MinE (1 μM). Scale bar is 50 μm, frames are 9 s apart.

Movie s9
Movie of a double spiral of Min proteins. MinD (1 μM), MinE (0.5 μM), doped with 10% Alexa647-labeled MinE. Scale bar is 50 μm, frames are 14 s apart.

Movie s10 (A)
Simulation of Min protein dynamics. The MinD is shown in green, MinE in red. This movie corresponds to Fig.4A in the article. The concentrations range for MinD from 0 to ~6 × 106 μm–2, and for MinE from 0 to ~5.4 × 106 μm-2. In total a square of side length 900 μm is shown, for a period of 14.5 min.

Movie s10 (B)
Extract of Movie S10 corresponding to the area shown in Fig.4A in the article. The square has a side length of 400 μm.

Movie s11
Simulation of Min protein dynamics. Only MinE is shown. This movie corresponds to Fig.4D in the article. The parameters are as in Movie S10 but different initial concentrations have been chosen. The concentration range equals the one in Movie S10. Shown is a square of side length 540 ?m for a period of 11.5 min.

Movie s12 (A)
Simulation of Min protein dynamics according to the Min-oscillation model presented in (5). Color coding is as in Movie S10. In total, a square of 900 mm has been simulated with periodic boundary conditions. Shown is a patch of 600 mm side length covering 4 min. The total concentration of MinD in the system was 1 mM, and of MinE 0.34 mM. The remaining parameters were s_de = 0.67s⁻¹, s_D = 0.17s⁻¹, s_dD = 2.6 · 10⁻⁷ mm²/s, s_E = 1.5 · 10⁻⁵ mm²/s, D_D = D_E = 60 mm²/s, c_max = 12.6 · 10⁶ mm⁻² . The exchange of ADP for ATP in unbound MinD was assumed to be instantaneous. The use of a finite exchange rate did not give qualitative changes. The concentrations range for MinD from 0 to ~12 · 10⁶ mm⁻², and for MinE from 0 to ~3.8 · 10⁶ mm⁻².

Movie s12 (B)
Simulation of the model presented in (5). In total, a square of 450 mm has been simulated with periodic boundary conditions. The movie covers 1 min. The total con­centration of MinD in the system was 1 mM, and of MinE 0.35 mM. The remaining parame­ters were s_de = 0.7 s⁻¹, s_{ADP →ATP} = 1 s⁻¹, s_D = 0.098 s⁻¹, s_dD = 6.6 · 10⁻⁷ mm²/s, s_E = 4 · 10⁻⁵ mm²/s, D_D = D_E = 63 mm²/s, c_max = 18.75 · 10⁶ mm⁻². The concentrations range for MinD from 0 to ~17.25 · 10⁶ mm⁻², and for MinE from 0 to ~7.5 · 10⁶ mm⁻².

Movie s13
Simulation of Min protein dynamics according to the Min-oscillation model pre­sented in (6). Color coding is as in Movie S10. Shown is a square of 250 mm side length covering 20 min simulated with periodic boundary conditions. The total concentration of MinD in the system was 1 mM, and for MinE 1 mM. The remaining parameters were s_de = 0.002 s⁻¹, s_D = 0.0004 s⁻¹, s_E = 3.3 · 10⁻⁹ mm²/s, c_max = 6 · 10⁶ mm⁻² . The concentrations range for MinD from 0 to ~1 · 10⁶ mm⁻², and for MinE from 0 to ~6 · 10⁵ mm⁻². The buffer proteins were assumed to be well-stirred, i.e., spatial variations of the concentrations within the buffer were neglected.

To view these movies, download a QuickTime viewer.