1. Volume 113, Issue 12, Pages 2621-2633 (December 2017)
Myoglobin and α-Lactalbumin Form Smaller Complexes with the Biosurfactant Rhamnolipid Than with SDS 
Henriette Gavlshøj Mortensen, Jens Kvist Madsen, Kell K. Andersen, Thomas Vosegaard, G. Roshan Deen, Daniel E. Otzen, Jan Skov Pedersen 
Biophysical Journal 
Volume 113, Issue 12, Pages (December 2017)
DOI: /j.bpj
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2. Figure 1
(a) Given here is the molecular structure of mono- and di-rhamnolipid. (b and c) Shown here is a simplified illustration of mono-RL. Different conformations lead to different results in electron densities of the core and shell, respectively, depending on the definition of head and tail. Here the sugar is drawn in dark shading, the oxygen-rich part of the fatty acid chains is in light shading, and the pure hydrocarbon chains are in solid representation.
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3. Figure 2 SLS data of 2 wt % RL.
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4. Figure 3
(a) Given here are SAXS data for various RL concentrations 7.6 mM (squares, black), 9.5 mM (circles, red), 12.7 mM (top triangles, green), and 20.3 mM (bottom triangles, blue) measured at 25°C with respective model fits (lines) for a prolate core-shell micelle. (Inset) Shown here is rescaled data normalized by concentration and demonstrating stable particle shape within the studied concentration range of RL. (b) Shown here is the p(r) function of 20.3 mM RL at 25°C obtained from IFT. Because no change in shape was observed within the studied concentration range, only one p(r) function is shown. To see this figure in color, go online.
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5. Figure 4
Plots of SAXS data for pure RL at 11 mM (solid line), pure αLA at 2.3 mg/mL (dashed line), a calculated linear combination of RL at 11 mM and αLA at 2.3 mg/mL(dashed-dot line), and experimental data of a αLA-RL mixture (dots).
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6. Figure 5
(a) Given here are SAXS data for 2.5 mg/mL αLA measured at 25°C mixed with 5.6 mM RL (bottom triangles, blue), 6.5 mM RL (top triangles, green), 9.2 mM RL (circles, red), and 11 mM RL (squares, black) with respective model fits (lines). Quality of the fits is χ2 = 6.8, 5.3, 4.2, and 4.3, respectively. A schematic drawing of the complex as described in the model is included in the figure. The hand-drawn line in the shell of the particle represents partly unfolded protein. (b) Shown here is scattering from pure αLA (bottom triangles, blue) and αLA with low RL concentrations of 0.7 mM (top triangles, green), 1.9 mM (circles, red), and 3.6 mM (squares, black). Pure αLA and αLA + 0.7 mM RL are fitted by the calculated scattering from PDB: 1F6R (lines) using the software CRYSOL. Quality of the fits for the data with pure αLA and 0.7 mM RL is χ2 = 1.7 and 2, respectively. (c) Given here are the p(r) functions obtained from IFT fitting of 2.5 mg/mL αLA with varying RL concentrations as indicated in the graph. (Dashed line in top graph) This shows the p(r) function for pure RL corresponding to 11 mM. To see this figure in color, go online.
Biophysical Journal  , DOI: ( /j.bpj )
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7. Figure 6
(a) Shown here are SAXS data for 2.5 mg/mL Mb measured at 25°C mixed with 4.7 mM RL (rhombus, light blue), 6.9 mM RL (bottom triangles, blue), 7.6 mM RL (top triangles, green), 9.3 mM RL (circles, red), and 11 mM (squares, black), with respective model fits (lines). Quality of the fits is χ2 = 2.4, 1.9, 1.4, 2.5, and 2.1, respectively. A schematic drawing of the complex as described in the model is included in the figure. The hand-drawn line in the shell of the particle represents partly unfolded protein. (b) Shown here is scattering from pure Mb (top triangles, green) and Mb with low RL concentrations of 0.7 mM (circles, red) and 2.7 mM (squares, black). Pure Mb and Mb + 0.7 mM RL were fitted by the calculated scattering from PDB: 1YMB (line) using CRYSOL. Quality of the fits for the data with pure Mb and 0.7 mM RL is χ = 1 and 1.1, respectively. (c) Given here are the p(r) functions obtained from IFT fitting from 2.5 mg/mL Mb with varying RL concentrations as indicated in the graph and measured at 25°C. (Dashed line in top graph) This shows the p(r) function for pure RL corresponding to 11 mM. To see this figure in color, go online.
Biophysical Journal  , DOI: ( /j.bpj )
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8. Figure 7
(a) Given here are SAXS data for SDS micelles at varying concentrations of 7.5 mM (rhombus, light blue), 8.7 mM (bottom triangles, blue), 9.5 mM (top triangles, green), 14.4 mM (circles, red), and 19.3 mM (squares, black) measured at 22°C. Oblate-shaped particle model fits are shown as lines. The upturn in intensity at low q is probably due to the sample being measured close to the Krafft temperature, where counterions associate with micelles, which may in turn lead to attractive interactions between the micelles. (Inset) Shown here are data normalized by concentration. The scattering data do not appear to be perfectly aligned, which we ascribe to the relatively high cmc as well as low concentrations, leading to limited data quality. Data modeling shows no signs of change of the particle shape within the studied concentration range of SDS. (b) Shown here is the p(r) function for 8.7 mM SDS. The negative dip indicates core-shell shape. Because no change in shape was observed within the studied concentration range, only one function is shown. To see this figure in color, go online.
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9. Figure 8
(a) Given here are SAXS data for 2.1 mg/mL αLA (squares, black) measured at 22°C mixed varying SDS concentrations of 0.8 mM (circles, red), 2 mM (top triangles, green), 4 mM (bottom triangles, blue), 6 mM (rhombus, light blue), 9 mM (left triangles, magenta), and 12 mM (right triangles, dark yellow) with respective model fits (lines). (Inset) Show here is scattering from pure αLA fitted by the calculated scattering from the PDB: 1F6R (line) using CRYSOL. (b) Given here are the p(r) functions obtained from IFT fitting from 2.1 mg/mL αLA with varying SDS concentrations as indicated in the graph, and measured at 22°C. (Dashed line in top graph) This shows the p(r) function for pure SDS corresponding to 12 mM. To see this figure in color, go online.
Biophysical Journal  , DOI: ( /j.bpj )
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10. Figure 9
(a) Given here are fitting parameters RC (square), ε (sphere), D (rhombus), and calculated parameter Nagg (triangle), describing the αLA-SDS complex as function of SDS concentration. Errors were <5% for ε (except for 4–6 mM with an error of 11%) and ∼1% for RC . No errors are given for Nagg because this is not a fitting parameter but instead a calculated output based on ε and RC. (b) Given here is the number of αLA per complex. The protein content is generally larger than in RL-containing complexes. (c) Given here is the number of SDS molecules in the complex per αLA calculated from SAXS fitting parameters (open square) (method shown in the Supporting Material) compared to results from ITC measurements by Otzen et al. (13) (circle).
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