Electronic Supplementary Material (ESI) for Molecular BioSystems.
This journal is © The Royal Society of Chemistry 2016
Folding Propensity of Intrinsically Disordered Proteins by Osmotic Stress
Supporting Information
Amanda L. Mansouri,a Laura N. Grese, a,b Erica L. Rowe, a,b James Pino,c S. Chakra
Chennubhotla,d Arvind Ramanathan,c Hugh M. O’Neill,b Valerie Berthelier,a and Christopher B.
Stanleyb*
a
Department of Medicine, Graduate School of Medicine, University of Tennessee Health Science
Center, Knoxville, Tennessee 37920
b
Biology and Soft Matter Division Oak Ridge National Laboratory, P.O. Box 2008, Oak Ridge,
Tennessee 37831
c
Health Data Sciences Institute, Computational Science and Engineering Division, Oak Ridge
National Laboratory, Oak Ridge, Tennessee 37831
d
Department of Computational and Systems Biology, University of Pittsburgh, Pennsylvania
15260
Corresponding Author
*
E-mail: [email protected]
1 A PPRSISPSALQDLLRTLKSPSSPQQQQQVLNILKSNPQLMAAFIKQRTAKYVANQPGMQ 59
------HHHHHHHHHHH-------HHHHHHHHH---HHHHHHHHHHHHHH--------- 59
Total Residues: H: 34
Percent: H: 57.6
E: 0
E: 0
T: 0
T: 0
B EQGPHGSQNRPLLRNSLDDLLGPPSNAEGQSDERALLDQLHTFLSNTDATGLEEIDRALGIPELVNQGQALESK 74
--------------------E-----------HHHHHHHHHHHHH---HHHHHHHHHH---HHHHH-------- 74
Total Residues: H: 28
Percent: H: 37.8
E: 1
E: 1.4
T: 0
T: 0
Figure S1. The JPred4 secondary structure predictions for (A) NCBD and (B) ACTR, where H =
helix, S = sheet, and T = turn.
2 5
A -1
[θ] (10 deg cm decimol )
15
-1
[θ] (10 deg cm decimol )
20
10
2
2
5
0
0
-5
-10
3
3
-5
B -10
-15
-20
-20
190
-15
200
210 220 230 240
wavelength (nm)
250
260
190
200
210 220 230 240
wavelength (nm)
250
260
Figure S2. CD spectra for NCBD with (A) EG added from 0 to 40% (v/v) and (B) TEG added
from 0 to 40% (v/v). The arrows indicate the CD spectral changes that occur with increasing
osmolyte concentration.
3 5
-1
[θ] (10 deg cm decimol )
0
B 0
-5
2
-5
2
-1
[θ] (10 deg cm decimol )
A -10
3
3
-10
-15
-15
-20
-20
190
200
210 220 230 240
wavelength (nm)
260
190
200
210 220 230 240
wavelength (nm)
250
260
C 0
-5
-10
3
2
-1
[θ] (10 deg cm decimol )
5
250
-15
-20
190
200
210 220 230 240
wavelength (nm)
250
260
Figure S3. CD spectra for ACTR with (A) EG added from 0 to 40% (v/v), (B) xylitol added
from 0 to 4 molal, and (C) TEG added from 0 to 40% (v/v). The arrows indicate the CD spectral
changes that occur with increasing osmolyte concentration. Spectra in orange are the two highest
osmolyte concentrations and that begin to show deviations from the 205 nm isodichroic point.
4 -3
-1
-13
[θ]222 (10 deg cm dmol )
EG
TEG
PEG 400
EG
EG (run 2)
TEG
xylitol
PEG 400
-4
2
-14
3
-15
3
2
-1
[θ]222 (10 deg cm dmol )
-12
-16
-17
A -18
0
2
4
6
8
[osmolyte] (Osm)
10
12
-5
-6
-7
B -8
0
2
4
6
8
[osmolyte] (Osm)
10
12
Figure S4. Change in CD mean residue ellipticity ([q]222) with osmolality for (A) NCBD and (B)
ACTR.
5 -4.0
A -1
ln[I(Q) (cm )]
-4.2
-4.4
-4.6
-4.8
0
2
4
6
2
8
10x10
-3
-2
Q (Å )
-4.0
B -1
ln[I(Q) (cm )]
-4.2
-4.4
-4.6
-4.8
0
2
4
6
2
8
10x10
-3
-2
Q (Å )
-4.0
C -1
ln[I(Q) (cm )]
-4.2
-4.4
-4.6
-4.8
0
2
4
6
2
-2
Q (Å )
8
10x10
-3
Figure S5. Guinier plots and fits (solid lines) of the SANS data for NCBD with (A) 0, (B) 1.63,
and (C) 3.67 osmolal d6-EG.
6 0.52
0.50
fh
0.48
0.46
0.44
0.42
0.40
0
2
4
6
8
[EG] (osm)
10
12
Figure S6. NCBD fraction helicity, fh, as a function of EG osmolality. Here, fh is calculated as
absolute using Equation 1 in the main text with [q]222,h = –36,000 deg cm2 dmol-1. The sigmoid fit
(solid line), f h = A (1+ exp[([EG]1/ 2 − [EG]) τ ]) + f h,0 yielded A = 0.13 ± 0.02, [EG]1/2 = 4.4 ±
0.5, t = 2.2 ± 0.6, and fh,0 = 0.38 ± 0.01. The fit was used to interpolate fh vs. [EG] for the Figure
5 inset.
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