Supplementary Materials for
The Energetics of Intermediates that Guide Polyproline Folding
Liuqing Shi, Alison E. Holliday, Matthew S. Glover, Michael A. Ewing, David H. Russell, and
David E. Clemmer*
*Corresponding Author. Email: clemmer@indiana.edu
This file includes:
Supplementary Text
Tables S1 to S8
Figs. S1 to S35
References
S1
Supplementary Text:
Determination of Arrhenius activation parameters. For the PPI→PPII transition, the
Arrhenius activation parameters presented here were determined using additional measurements
together with our prior work (1) and represent a more comprehensive assessment of the kinetics
data and the associated error analysis. As is shown in Table S3, the obtained Arrhenius
parameters display similar values to the published values (1). Although the value of ∆𝑆 ǂ for the
E→H step (−7 ± 21 kJ·mol−1) is negative in our more thorough analysis, given the high standard
deviation associated with this number, the value documented here still agrees with our published
work (14 kJ·mol−1) (1). This finding indicates that the entropic barrier for the E→H step is very
low or absent.
S2
Table S1. Summary of the Rate Constants (in × 10−5 s-1) of Each Step in the Proposed Mechanism
for the Five Explored Temperatures of PPI→PPII transition.a,b
rate
5 °C
15 °C
23 °C
35 °C
45 °C
constant
kAB
33 ± 4.6
129 ± 13
207 ± 18
761 ± 30
1911 ± 275
kBC
7.4 ± 1.0
24 ± 2.1
40 ± 2.1
194 ± 7.7
498 ± 18
kCD
16 ± 2.2
51 ± 7.4
84 ± 6.7
453 ± 28
1306 ± 71
kDE
15 ± 4.4
71 ± 9.2
83 ± 3.6
487 ± 32
1640 ± 105
kEF
23 ± 4.1
68 ± 2.2
106 ± 4.8
463 ± 30
115 ± 29
kEG
15 ± 0.2
29 ± 1.8
61 ± 4.9
244 ± 24
649 ± 82
kEH
0.5 ± 0.2
4.6 ± 1.6
3.3 ± 0.7
34 ± 1.4
116 ± 23
a
The average and standard deviation were obtained from four independent measurements.
b
The proposed PPI→PPII transition mechanism is shown in Fig. 3, top left.
i
S3
Table S2. Summary of the Rate Constants (in × 10−5 s-1) of Each Step in the Proposed Mechanism
for the Five Explored Temperatures of the PPII→PPI transition.a,b
rate
5 °C
15 °C
23 °C
35 °C
45 °C
constant
kFD*
0.03 ± 0.00
0.17 ± 0.08
0.18 ± 0.02
1.4 ± 0.2
2.8 ± 0.8
kFE*
0.06 ± 0.00
0.18 ± 0.07
0.33 ± 0.03
1.5 ± 0.4
3.7 ± 1.1
kFG*
0.17 ± 0.02
0.42 ± 0.03
2.0 ± 0.1
9.1 ± 0.9
19 ± 2.9
kFH*
0.02 ± 0.00
0.03 ± 0.01
0.16 ± 0.0
0.9 ± 0.6
1.4 ± 0.4
kG*D*
0.15 ± 0.07
0.33 ± 0.19
2.9 ± 0.4
25 ± 8.5
54 ± 6.7
kG*E*
0.52 ± 0.03
1.5 ± 0.1
7.0 ± 0.5
21 ± 5.2
23 ± 6.8
kH*D*
0.13 ± 0.04
0.13 ± 0.03
1.8 ± 0.2
11 ± 10
43 ± 7.1
kH*E*
0.30 ± 0.03
0.08 ± 0.04
3.7 ± 0.4
17 ± 9.9
5.6 ± 0.8
i
kD*C*
0.82 ± 0.36
2.8 ± 1.3
15 ± 1.6
130 ± 12
314 ± 23
kE*C*
1.2 ± 0.1
3.6 ± 0.7
14 ± 0.8
59 ± 13
94 ± 6.9
kC*A
1.4 ± 0.1
2.4 ± 0.5
5.0 ± 0.3
17 ± 2.2
50 ± 4.1
kC*B
0.77 ± 0.05
1.6 ± 0.3
2.9 ± 0.6
11 ± 1.9
24 ± 3.1
ai
The average and standard deviation were obtained from four independent measurements.
b
The proposed PPII→PPI transition mechanism is shown in Fig. 3, top right.
S4
Table S3. Arrhenius activation energy, 𝐸𝑎 , preexponential factor, A, enthalpy of activation, ∆𝐻 ǂ ,
entropy of activation, ∆𝑆 ǂ , and Gibbs free energy of activation, ∆𝐺 ǂ , for each transition step in the
proposed mechanism for the PPI→PPII transition.
Transition
𝐸𝑎 ,b,c
∆𝐻 ǂ , b,d
∆𝑆 ǂ , b,c,e
∆𝐺 ǂ , b,f
b,c −1
A,
s
stepa
kJ·mol−1
kJ·mol−1 J·mol−1·K−1
kJ·mol−1
A→B
73 ± 2(74 ± 4)
1010.3 ± 0.3(1010.4 ± 0.7) 71 ± 2 −57 ± 6 (−49)
87 ± 3
B→C
78 ± 2(78 ± 2)
1010.4 ± 0.3(1010.6 ± 0.4) 75 ± 2 −53 ± 7 (−49)
91 ± 3
11.4 ± 0.4
11.5 ± 0.3
C→D
81 ± 2(81 ± 2)
10
(10
) 79 ± 2 −36 ± 8 (−33)
89 ± 4
11.9 ± 0.6
12.1 ± 1.2
D→E
84 ± 4(85 ± 7)
10
(10
) 82 ± 4 −24 ± 12(−9)
89 ± 5
E→F
72 ± 2(72 ± 3)
109.9 ± 0.4 (109.9 ± 0.5)
70 ± 2 −65 ± 7 (−61)
89 ± 3
i
9.5 ± 0.4
9.5 ± 0.5
E→G
71 ± 2(71 ± 3)
10
(10
)
69 ± 2 −72 ± 8 (−69)
90 ± 3
E→H
97 ± 6(95 ± 11) 1012.9 ± 1.1(1012.6 ± 1.9) 94 ± 6
−7 ± 21(14)
96 ± 9
a
The transition steps are associated with the transition mechanism proposed in Fig. 3, top left.
bi
Uncertainties correspond to one deviation about the mean for four independent measurements.
c
The values shown in parentheses come from ref 1.
d
Values for the enthalpy of activation are calculated at 296.15 K using eq. 5.
e
Values for the entropy of activation are calculated at 296.15 K using eq. 6.
fs
Values for the Gibbs free energy of activation are calculated at 296.15 K using eq. 7.
S5
Table S4. Arrhenius activation energy, 𝐸𝑎 , preexponential factor, A, the enthalpy of activation,
∆𝐻 ǂ , the entropy of activation, ∆𝑆 ǂ , and the Gibbs free energy of activation, ∆𝐺 ǂ , for each transition
step in the proposed mechanism for the PPII→PPI transition.
Transition
𝐸𝑎 ,b
∆𝐻 ǂ , b,c
∆𝑆 ǂ , b,d
∆𝐺 ǂ , b,e
b −1
A,
s
stepa
kJ·mol−1
kJ·mol−1
J·mol−1·K−1
kJ·mol−1
9.4 ± 0.9
F→D*
85 ± 5
10
82 ± 5
−73 ± 17
104 ± 7
F→E*
77 ± 3
108.2 ± 0.6
75 ± 3
−96 ± 11
103 ± 4
11.5 ± 0.5
F→G*
92 ± 3
10
90 ± 3
−34 ± 10
100 ± 4
F→H*
83 ± 6
108.7 ± 1.0
80 ± 6
−86 ± 20
106 ± 8
17.0 ± 1.3
G*→D*
122 ± 7
10
120 ± 7
72 ± 24
99 ± 10
8.9 ± 0.8
G*→E*
75 ± 5
10
73 ± 5
−83 ± 16
97 ± 7
H*→D*
115 ± 10
1015.3 ± 1.8
112 ± 10
40 ± 34
100 ± 14
H*→E*
65 ± 9
106.7 ± 1.6
62 ± 9
−125 ± 31
99 ± 13
D*→C*
118 ± 5
1017.0 ± 0.8
116 ± 5
72 ± 16
94 ± 7
10.9 ± 0.6
E*→C*
84 ± 3
10
82 ± 3
−44 ± 11
95 ± 4
7.6 ± 0.5
C*→A
67 ± 3
10
64 ± 3
−108 ± 10
96 ± 4
C*→B
65 ± 2
107.0 ± 0.4
63 ± 2
−118 ± 8
98 ± 4
a
The transition steps are associated with the transition mechanism proposed in Fig. 3, top right.
b
Uncertainties correspond to one deviation about the mean for four independent measurements.
c
Values for the enthalpy of activation are calculated at 296.15 K using eq. 5.
d
Values for the entropy of activation are calculated at 296.15 K using eq. 6.
e
Values for the Gibbs free energy of activation are calculated at 296.15 K using eq. 7.
S6
Table S5. The enthalpy, ∆𝐻, the entropy, ∆𝑆, and the Gibbs free energy, ∆𝐺, of each conformer
for the PPI→PPII transition.a
Conformer
∆𝐻, c
∆𝑆, c
∆𝐺,c
typeb
kJ·mol−1
J·mol−1·K−1
kJ·mol−1
B
−22 ± 12
−81 ± 39
2 ± 17
C
−24 ± 10
−82 ± 33
0 ± 14
D
14 ± 12
69 ± 38
−6 ± 16
E
18 ± 6
98 ± 20
−11 ± 8
F
21 ± 9
130 ± 30
−18 ± 13
G
20 ± 10
118 ± 33
−15 ± 14
H
25 ± 10
126 ± 34
−12 ± 14
a
∆𝐻, ∆𝑆, and ∆𝐺 associated with each transition step are determined by subtracting the value of
the precursor from that of the product.
b
The conformer types are assigned based on the CCS distributions shown in Fig. S34.
c
Uncertainties correspond to one deviation about the mean for three independent measurements
when conformer A is considered as the initial conformer.
S7
Table S6. The enthalpy, ∆𝐻, the entropy, ∆𝑆, and the Gibbs free energy, ∆𝐺, of each conformer
for the PPII→PPI transition.ai
Conformer
∆𝐻,c
∆𝑆, c
∆𝐺,c
typeb
kJ·mol−1
J·mol−1·K−1
kJ·mol−1
A
56 ± 10
235 ± 31
−14 ± 13
B*
70 ± 9
276 ± 29
−12 ± 12
C*
96 ± 15
342 ± 49
−5 ± 21
D*
62 ± 10
203 ± 33
2 ± 14
E*
51 ± 11
172 ± 36
0 ± 15
G*
43 ± 9
144 ± 28
0 ± 12
H*
41 ± 10
127 ± 33
3 ± 14
a
∆𝐻, ∆𝑆, and ∆𝐺 associated with each transition step are determined by subtracting the value of
the precursor from that of the product.
bi
The conformer types are assigned based on the CCS distributions shown in Fig. S35.
c
Uncertainties correspond to one deviation about the mean for three independent measurements
when conformer F is considered as the initial conformer.
S8
Table S7. The residual sum of squares from fitting each of the eight conformers (A-H) in the 45 °C
abundance profiles using 13 candidate PPI→PPII transition fit models.a
a
bi
residual sum of squares from fitting (× 10−3)
Fit
modelb
A
B
C
D
E
F
G
H
F1
F2
F3
F4
F5
F6
F7
F8
F9
F10
F11
F12
F13
1.190
1.190
1.190
1.190
1.190
1.190
1.200
1.190
1.200
1.200
1.190
1.190
1.190
2.060
58.80
58.80
54.93
54.93
74.66
2.170
2.060
2.170
2.170
2.060
2.060
2.060
0.967
7.270
7.270
17.78
17.78
12.84
2.880
2.880
4.310
2.880
0.967
0.967
0.967
6.500
18.16
18.16
11.53
11.53
8.040
58.94
58.94
68.28
66.53
3.940
3.230
6.500
2.310
4.210
1.670
9.440
9.440
9.350
86.99
86.99
77.30
91.93
2.940
5.150
2.310
1.710
15.00
64.83
37.57
34.34
28.15
1253
56.03
871.6
1296
100.4
53.97
124.6
1.250
4.700
78.83
179.4
14.93
254.7
1897
78.38
1981
1053
109.2
19.77
139.9
1.060
1.060
13.54
0.803
2.090
0.781
18.81
7.680
14.56
136.3
2.390
18.23
1.220
Sum for all
conformers
17.05
110.4
244.3
312.6
146.2
389.7
3322
294.2
3020
2650
223.1
104.6
278.8
The conformer types are assigned based on the CCS distributions shown in Fig. 2.
The fitting models are associated with the transition routes shown in Figs. S8 to S20.
S9
Table S8. The residual sum of squares from fitting each of the eight conformers (A-H*) in the
45 °C abundance profiles using 12 candidate PPII→PPI transition fit models.a
a
bi
residual sum of squares from fitting (× 10−3)
Fit
modelb
A
B*
C*
D*
E*
F
G*
H*
R1
R2
R3
R4
R5
R6
R7
R8
R9
R10
R11
R12
4.910
6.070
5.010
4.730
7.630
8.490
16.17
250.0
131.5
131.5
418.5
261.7
2.230
3.330
2.190
2.220
4.240
8.930
23.57
229.0
228.7
228.7
92.32
338.0
7.170
7.870
6.690
7.180
6.290
5.050
30.96
72.60
29.46
35.10
35.10
19.55
1.120
1.110
1.130
1.100
1.180
1.170
1.180
1.190
1.220
1.200
1.200
34.22
0.961
1.000
0.976
1.010
1.200
1.150
1.200
1.250
1.150
1.290
1.290
1.240
9.010
9.590
9.590
9.590
7.990
7.990
7.990
7.990
9.590
9.590
9.590
9.590
1.150
1.270
1.270
1.270
1.600
1.600
1.600
1.600
1.920
1.920
1.920
2.640
0.633
0.632
0.632
0.632
0.484
0.484
0.484
0.484
0.551
0.551
0.551
0.390
Sum for all
conformers
27.18
30.87
27.49
27.73
30.61
34.86
83.15
564.1
404.1
409.9
560.5
667.3
The conformer types are assigned based on the CCS distributions shown in Fig. 2.
The fitting models are associated with the transition routes shown in Figs. S21 to S32.
S10
H* G* F E*D* C* B*
A
5 C
298 hr
231 hr
Normalized Intensity
163 hr
122 hr
97 hr
26 hr
1 hr
H2O
250
270
290
310
330
350
CCS (Å2)
Fig. S1. Collision cross section distributions for the [Pro13+2H]2+ ions obtained at different transition times, showing
the transition from PPII→PPI in 97:1:2 1-propanol:H2O:HOAc (v:v:v) at 5 °C. In order to illustrate the initial distribution
for the transition, the distribution for the ions formed by electrospraying H 2O is also shown at the bottom. The transition
times when the distributions were obtained are indicated in each trace, and dashed lines delineate the collision cross
section region for each conformer type.
S11
Relative Abundance
1.0
D*
5 C
0.8
A
C*
E*
F
B*
G*
0.6
H*
0.4
0.2
0.0
0
50
100
150
200
250
300
Time (hr)
Fig. S2. Relative abundance of different conformer types for the transition from PPII→PPI in 97:1:2 1propanol:H2O:HOAc (v:v:v) at 5 °C as a function of transition time, and the transition pathway used to fit the data
points. The lines show the best fitting results obtained on the basis of the transition mechanism shown here. Various
colors are used to represent different conformations: black is conformer A, red is conformer B*, blue is conformer C*,
magenta is conformer D*, green is conformer E*, orange is conformer F, violet is conformer G*, and olive is
conformer H*.
S12
H* G* F E*D* C* B*
A
15 C
138 hr
118 hr
92 hr
Normalized Intensity
64 hr
46 hr
27 hr
12 hr
4 hr
H2O
250
270
290
310
330
350
CCS (Å2)
Fig. S3. Collision cross section distributions for the [Pro13+2H]2+ ions obtained at different transition times, showing
the transition from PPII→PPI in 97:1:2 1-propanol:H2O:HOAc (v:v:v) at 15 °C. In order to illustrate the initial
distribution for the transition, the distribution for the ions formed by electrospraying H 2O is also shown at the bottom.
The transition times when the distributions were obtained are indicated in each trace, and dashed lines delineate the
collision cross section region for each conformer type.
S13
Relative Abundance
1.0
D*
15 C
0.8
E*
F
A
C*
B*
G*
0.6
H*
0.4
0.2
0.0
0
25
50
75
100
125
Time (hr)
Fig. S4. Relative abundance of different conformer types for the transition from PPII→PPI in 97:1:2 1propanol:H2O:HOAc (v:v:v) at 15 °C as a function of transition time, and the transition pathway used to fit the data
points. The lines show the best fitting results obtained on the basis of the transition mechanism shown here. Various
colors are used to represent different conformations: black is conformer A, red is conformer B*, blue is conformer C*,
magenta is conformer D*, green is conformer E*, orange is conformer F, violet is conformer G*, and olive is
conformer H*.
S14
H* G* F E*D* C* B*
A
35 C
479 min
372 min
Normalized Intensity
256 min
187 min
135 min
79 min
27 min
8 min
H2O
250
270
290
310
330
350
CCS (Å2)
Fig. S5. Collision cross section distributions for the [Pro13+2H]2+ ions obtained at different transition times, showing
the transition from PPII→PPI in 97:1:2 1-propanol:H2O:HOAc (v:v:v) at 35 °C. In order to illustrate the initial
distribution for the transition, the distribution for the ions formed by electrospraying H 2O is also shown at the bottom.
The transition times when the distributions were obtained are indicated in each trace, and dashed lines delineate the
collision cross section region for each conformer type.
S15
Relative Abundance
1.0
D*
35 C
0.8
E*
F
A
C*
B*
G*
0.6
H*
0.4
0.2
0.0
0
80
160
240
320
400
480
Time (min)
Fig. S6. Relative abundance of different conformer types for the transition from PPII→PPI in 97:1:2 1propanol:H2O:HOAc (v:v:v) at 35 °C as a function of transition time, and the transition pathway used to fit the data
points. The lines show the best fitting results obtained on the basis of the transition mechanism shown here. Various
colors are used to represent different conformations: black is conformer A, red is conformer B*, blue is conformer C*,
magenta is conformer D*, green is conformer E*, orange is conformer F, violet is conformer G*, and olive is
conformer H*.
S16
H* G* F E*D* C* B*
A
45 C
210 min
175 min
Normalized Intensity
130 min
81 min
54 min
35 min
13 min
2 min
H2O
250
270
290
310
330
350
CCS (Å2)
Fig. S7. Collision cross section distributions for the [Pro13+2H]2+ ions obtained at different transition times, showing
the transition from PPII→PPI in 97:1:2 1-propanol:H2O:HOAc (v:v:v) at 45 °C. In order to illustrate the initial
distribution for the transition, the distribution for the ions formed by electrospraying H2O is also shown at the bottom.
The transition times when the distributions were obtained are indicated in each trace, and dashed lines delineate the
collision cross section region for each conformer type.
S17
Relative Abundance
1.0
F1
0.8
G
A
B
C
D
E
F
H
0.6
0.4
0.2
0.0
0
2
4
6
8
10
12
14
16
Time (min)
Fig. S8. Relative abundance of different conformer types for the PPI→PPII transition in 10:88:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and the proposed transition route (F1) that
works for fitting the data points. Various colors are used to represent different conformations: black is conformer A,
red is conformer B, blue is conformer C, magenta is conformer D, green is conformer E, orange is conformer F,
violet is conformer G, and olive is conformer H. Fitting statistics are provided in Table S7, and the lines shown here
represent the best fitting results. The mathematical functions used to fit the data points were created by solving a
series of differential equations that describe the reaction rate of each conformer involved in the transition route:
𝑦 ′ (A) = −𝑘𝐴 × 𝑦(𝐴) , 𝑦 ′ (𝐵) = 𝑘𝐴 × 𝑦(𝐴) − 𝑘𝐵 × 𝑦(𝐵) , 𝑦 ′ (𝐶) = 𝑘𝐵 × 𝑦(𝐵) − 𝑘𝐶 × 𝑦(𝐶) , 𝑦 ′ (𝐷) = 𝑘𝐶 × 𝑦(𝐶) − 𝑘𝐷 × 𝑦(𝐷) , 𝑦 ′ (𝐸) =
𝑘𝐷 × 𝑦(𝐷) − 𝑘𝐸 × 𝑦(𝐸) , 𝑦 ′ (F) = −𝑘𝐸𝐹 × 𝑦(𝐸) , 𝑦 ′ (G) = −𝑘𝐸𝐺 × 𝑦(𝐸) , 𝑦 ′ (H) = −𝑘𝐸𝐻 × 𝑦(𝐸) .
S18
Relative Abundance
1.0
F2
G
B
0.8
D
A
E
F
C
0.6
H
0.4
0.2
0.0
0
2
4
6
8
10
12
14
16
Time (min)
Fig. S9. Relative abundance of different conformer types for the PPI→PPII transition in 10:88:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (F2) that does not
work for fitting the data points. Various colors are used to represent different conformations: black is conformer A,
red is conformer B, blue is conformer C, magenta is conformer D, green is conformer E, orange is conformer F,
violet is conformer G, and olive is conformer H. Fitting statistics are provided in Table S7, and the lines shown here
represent the best fitting results. The mathematical functions used to fit the data points were created by solving a
series of differential equations that describe the reaction rate of each conformer involved in the transition route:
𝑦 ′ (A) = −𝑘𝐴 × 𝑦(A) , 𝑦 ′ (𝐵) = 𝑘𝐴𝐵 × 𝑦(A) − 𝑘𝐵 × 𝑦(𝐵) , 𝑦 ′ (𝐶) = 𝑘𝐴𝐶 × 𝑦(𝐴) − 𝑘𝐶 × 𝑦(𝐶) , 𝑦 ′ (𝐷) = 𝑘𝐵 × 𝑦(𝐵) + 𝑘𝐶 × 𝑦(𝐶) −
𝑘𝐷 × 𝑦(𝐷) , 𝑦 ′ (𝐸) = 𝑘𝐷 × 𝑦(𝐷) − 𝑘𝐸 × 𝑦(𝐸) , 𝑦 ′ (F) = −𝑘𝐸𝐹 × 𝑦(𝐸) , 𝑦 ′ (G) = −𝑘𝐸𝐺 × 𝑦(𝐸) , 𝑦 ′ (H) = −𝑘𝐸𝐻 × 𝑦(𝐸) .
S19
Relative Abundance
1.0
F3
B
E
D
0.8 A
C
0.6
F
H
G
0.4
0.2
0.0
0
2
4
6
8
10
12
14
16
Time (min)
Fig. S10. Relative abundance of different conformer types for the PPI→PPII transition in 10:88:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (F3) that does not work
for fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B, blue is conformer C, magenta is conformer D, green is conformer E, orange is conformer F, violet is
conformer G, and olive is conformer H. Fitting statistics are provided in Table S7, and the lines shown here represent
the best fitting results. The mathematical functions used to fit the data points were created by solving a series of
differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (A) = −𝑘𝐴 ×
𝑦(A) , 𝑦 ′ (𝐵) = 𝑘𝐴𝐵 × 𝑦(A) − 𝑘𝐵 × 𝑦(𝐵) , 𝑦 ′ (𝐶) = 𝑘𝐴𝐶 × 𝑦(𝐴) − 𝑘𝐶 × 𝑦(𝐶) , 𝑦 ′ (𝐷) = 𝑘𝐵 × 𝑦(𝐵) + 𝑘𝐶 × 𝑦(𝐶) − 𝑘𝐷 ×
𝑦(𝐷) , 𝑦 ′ (𝐸) = 𝑘𝐷𝐸 × 𝑦(𝐷) − 𝑘𝐸 × 𝑦(𝐸) , 𝑦 ′ (𝐺) = 𝑘𝐷𝐺 × 𝑦(𝐷) − 𝑘𝐺 × 𝑦(𝐺) , 𝑦 ′ (𝐻) = 𝑘𝐸 × 𝑦(𝐸) + 𝑘𝐺 × 𝑦(𝐺) − 𝑘𝐻 × 𝑦(𝐻) ,
𝑦 ′ (𝐹) = 𝑘𝐻 × 𝑦(𝐻) .
S20
Relative Abundance
1.0
F4
0.8
B
F
C
A
E
G
D
0.6
H
0.4
0.2
0.0
0
2
4
6
8
10
12
14
16
Time (min)
Fig. S11. Relative abundance of different conformer types for the PPI→PPII transition in 10:88:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (F4) that does not work
for fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B, blue is conformer C, magenta is conformer D, green is conformer E, orange is conformer F, violet is
conformer G, and olive is conformer H. Fitting statistics are provided in Table S7, and the lines shown here represent
the best fitting results. The mathematical functions used to fit the data points were created by solving a series of
differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (A) = −𝑘𝐴 ×
𝑦(A) , 𝑦 ′ (𝐵) = 𝑘𝐴𝐵 × 𝑦(A) − 𝑘𝐵 × 𝑦(𝐵) , 𝑦 ′ (𝐶) = 𝑘𝐴𝐶 × 𝑦(𝐴) − 𝑘𝐶 × 𝑦(𝐶) , 𝑦 ′ (𝐷) = 𝑘𝐴𝐷 × 𝑦(𝐴) − 𝑘𝐷 × 𝑦(𝐷) , 𝑦 ′ (𝐸) = 𝑘𝐵 × 𝑦(𝐵) +
𝑘𝐶 × 𝑦(𝐶) + 𝑘𝐷 × 𝑦(𝐷) − 𝑘𝐸 × 𝑦(𝐸) , 𝑦 ′ (𝐺) = 𝑘𝐸 × 𝑦(𝐸) − 𝑘𝐺 × 𝑦(𝐺) , 𝑦 ′ (𝐹) = 𝑘𝐺𝐹 × 𝑦(𝐺) , 𝑦 ′ (𝐻) = 𝑘𝐺𝐻 × 𝑦(𝐺) .
S21
Relative Abundance
1.0
F5
G
B
0.8
C
A
E
D
0.6
F
H
0.4
0.2
0.0
0
2
4
6
8
10
12
14
16
Time (min)
Fig. S12. Relative abundance of different conformer types for the PPI→PPII transition in 10:88:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (F5) that does not work
for fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B, blue is conformer C, magenta is conformer D, green is conformer E, orange is conformer F, violet is
conformer G, and olive is conformer H. Fitting statistics are provided in Table S7, and the lines shown here represent
the best fitting results. The mathematical functions used to fit the data points were created by solving a series of
differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (A) = −𝑘𝐴 ×
𝑦(A) , 𝑦 ′ (𝐵) = 𝑘𝐴𝐵 × 𝑦(A) − 𝑘𝐵 × 𝑦(𝐵) , 𝑦 ′ (𝐶) = 𝑘𝐴𝐶 × 𝑦(𝐴) − 𝑘𝐶 × 𝑦(𝐶) , 𝑦 ′ (𝐷) = 𝑘𝐴𝐷 × 𝑦(𝐴) − 𝑘𝐷 × 𝑦(𝐷) , 𝑦 ′ (𝐸) = 𝑘𝐵 × 𝑦(𝐵) +
𝑘𝐶 × 𝑦(𝐶) + 𝑘𝐷 × 𝑦(𝐷) − 𝑘𝐸 × 𝑦(𝐸) , 𝑦 ′ (𝐺) = 𝑘𝐸𝐺 × 𝑦(𝐸) , 𝑦 ′ (𝐹) = 𝑘𝐸𝐹 × 𝑦(𝐸) , 𝑦 ′ (𝐻) = 𝑘𝐸𝐻 × 𝑦(𝐸) .
S22
Relative Abundance
1.0
0.8
F6
B
A
C
F
G
D
H
0.6
E
0.4
0.2
0.0
0
2
4
6
8
10
12
14
16
Time (min)
Fig. S13. Relative abundance of different conformer types for the PPI→PPII transition in 10:88:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (F6) that does not work
for fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B, blue is conformer C, magenta is conformer D, green is conformer E, orange is conformer F, violet is
conformer G, and olive is conformer H. Fitting statistics are provided in Table S7, and the lines shown here represent
the best fitting results. The mathematical functions used to fit the data points were created by solving a series of
differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (A) = −𝑘𝐴 ×
𝑦(A) , 𝑦 ′ (𝐵) = 𝑘𝐴𝐵 × 𝑦(A) − 𝑘𝐵 × 𝑦(𝐵) , 𝑦 ′ (𝐶) = 𝑘𝐴𝐶 × 𝑦(𝐴) − 𝑘𝐶 × 𝑦(𝐶) , 𝑦 ′ (𝐷) = 𝑘𝐴𝐷 × 𝑦(𝐴) − 𝑘𝐷 × 𝑦(𝐷) , 𝑦 ′ (𝐸) = 𝑘𝐴𝐸 × 𝑦(𝐴) −
𝑘𝐸 × 𝑦(𝐸) , 𝑦 ′ (𝐺) = 𝑘𝐵 × 𝑦(𝐵) + 𝑘𝐶 × 𝑦(𝐶) + 𝑘𝐷 × 𝑦(𝐷) + 𝑘𝐸 × 𝑦(𝐸) − 𝑘𝐺 × 𝑦(𝐺) , 𝑦 ′ (𝐹) = 𝑘𝐺𝐹 × 𝑦(𝐺) , 𝑦 ′ (𝐻) = 𝑘𝐺𝐻 × 𝑦(𝐺) .
S23
Relative Abundance
1.0
F7
F
C
0.8 A
E
B
G
H
D
0.6
0.4
0.2
0.0
0
2
4
6
8
10
12
14
16
Time (min)
Fig. S14. Relative abundance of different conformer types for the PPI→PPII transition in 10:88:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (F7) that does not work
for fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B, blue is conformer C, magenta is conformer D, green is conformer E, orange is conformer F, violet is
conformer G, and olive is conformer H. Fitting statistics are provided in Table S7, and the lines shown here represent
the best fitting results. The mathematical functions used to fit the data points were created by solving a series of
differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (A) = −𝑘𝐴 ×
𝑦(A) , 𝑦 ′ (𝐵) = 𝑘𝐴 × 𝑦(A) − 𝑘𝐵 × 𝑦(𝐵) , 𝑦 ′ (𝐶) = 𝑘𝐵𝐶 × 𝑦(𝐵) − 𝑘𝐶 × 𝑦(𝐶) , 𝑦 ′ (𝐷) = 𝑘𝐵𝐷 × 𝑦(𝐵) − 𝑘𝐷 × 𝑦(𝐷) , 𝑦 ′ (𝐸) = 𝑘𝐶 × 𝑦(𝐶) +
𝑘𝐷 × 𝑦(𝐷) − 𝑘𝐸 × 𝑦(𝐸) , 𝑦 ′ (𝐺) = 𝑘𝐸 × 𝑦(𝐸) − 𝑘𝐺 × 𝑦(𝐺) , 𝑦 ′ (𝐹) = 𝑘𝐺𝐹 × 𝑦(𝐺) , 𝑦 ′ (𝐻) = 𝑘𝐺𝐻 × 𝑦(𝐺) .
S24
Relative Abundance
1.0
F8
0.8
G
C
A
E
B
F
D
0.6
H
0.4
0.2
0.0
0
2
4
6
8
10
12
14
16
Time (min)
Fig. S15. Relative abundance of different conformer types for the PPI→PPII transition in 10:88:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (F8) that does not work
for fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B, blue is conformer C, magenta is conformer D, green is conformer E, orange is conformer F, violet is
conformer G, and olive is conformer H. Fitting statistics are provided in Table S7, and the lines shown here represent
the best fitting results. The mathematical functions used to fit the data points were created by solving a series of
differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (A) = −𝑘𝐴 ×
𝑦(A) , 𝑦 ′ (𝐵) = 𝑘𝐴 × 𝑦(A) − 𝑘𝐵 × 𝑦(𝐵) , 𝑦 ′ (𝐶) = 𝑘𝐵𝐶 × 𝑦(𝐵) − 𝑘𝐶 × 𝑦(𝐶) , 𝑦 ′ (𝐷) = 𝑘𝐵𝐷 × 𝑦(𝐵) − 𝑘𝐷 × 𝑦(𝐷) , 𝑦 ′ (𝐸) = 𝑘𝐶 × 𝑦(𝐶) +
𝑘𝐷 × 𝑦(𝐷) − 𝑘𝐸 × 𝑦(𝐸) , 𝑦 ′ (𝐺) = 𝑘𝐸𝐺 × 𝑦(𝐸) , 𝑦 ′ (𝐹) = 𝑘𝐸𝐹 × 𝑦(𝐸) , 𝑦 ′ (𝐻) = 𝑘𝐸𝐻 × 𝑦(𝐸) .
S25
Relative Abundance
1.0
F9
0.8
F
C
A
D
B
G
H
E
0.6
0.4
0.2
0.0
0
2
4
6
8
10
12
14
16
Time (min)
Fig. S16. Relative abundance of different conformer types for the PPI→PPII transition in 10:88:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (F9) that does not work
for fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B, blue is conformer C, magenta is conformer D, green is conformer E, orange is conformer F, violet is
conformer G, and olive is conformer H. Fitting statistics are provided in Table S7, and the lines shown here represent
the best fitting results. The mathematical functions used to fit the data points were created by solving a series of
differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (A) = −𝑘𝐴 ×
𝑦(A) , 𝑦 ′ (𝐵) = 𝑘𝐴 × 𝑦(A) − 𝑘𝐵 × 𝑦(𝐵) , 𝑦 ′ (𝐶) = 𝑘𝐵𝐶 × 𝑦(𝐵) − 𝑘𝐶 × 𝑦(𝐶) , 𝑦 ′ (𝐷) = 𝑘𝐵𝐷 × 𝑦(𝐵) − 𝑘𝐷 × 𝑦(𝐷) , 𝑦 ′ (𝐸) = 𝑘𝐵𝐸 × 𝑦(𝐵) −
𝑘𝐸 × 𝑦(𝐸) , 𝑦 ′ (𝐺) = 𝑘𝐶 × 𝑦(𝐶) + 𝑘𝐷 × 𝑦(𝐷) + 𝑘𝐸 × 𝑦(𝐸) − 𝑘𝐺 × 𝑦(𝐺) , 𝑦 ′ (𝐹) = 𝑘𝐺𝐹 × 𝑦(𝐺) , 𝑦 ′ (𝐻) = 𝑘𝐺𝐻 × 𝑦(𝐺) .
S26
Relative Abundance
1.0
C
F10
0.8 A
E
B
F
H
D
G
0.6
0.4
0.2
0.0
0
2
4
6
8
10
12
14
16
Time (min)
Fig. S17. Relative abundance of different conformer types for the PPI→PPII transition in 10:88:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (F10) that does not work
for fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B, blue is conformer C, magenta is conformer D, green is conformer E, orange is conformer F, violet is
conformer G, and olive is conformer H. Fitting statistics are provided in Table S7, and the lines shown here represent
the best fitting results. The mathematical functions used to fit the data points were created by solving a series of
differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (A) = −𝑘𝐴 ×
𝑦(A) , 𝑦 ′ (𝐵) = 𝑘𝐴 × 𝑦(A) − 𝑘𝐵 × 𝑦(𝐵) , 𝑦 ′ (𝐶) = 𝑘𝐵𝐶 × 𝑦(𝐵) − 𝑘𝐶 × 𝑦(𝐶) , 𝑦 ′ (𝐷) = 𝑘𝐵𝐷 × 𝑦(𝐵) − 𝑘𝐷 × 𝑦(𝐷) , 𝑦 ′ (𝐸) = 𝑘𝐶 × 𝑦(𝐶) −
𝑘𝐸 × 𝑦(𝐸) , 𝑦 ′ (𝐺) = 𝑘𝐷 × 𝑦(𝐷) − 𝑘𝐺 × 𝑦(𝐺) , 𝑦 ′ (𝐻) = 𝑘𝐸 × 𝑦(𝐸) + 𝑘𝐺 × 𝑦(𝐺) − 𝑘𝐻 × 𝑦(𝐻) , 𝑦 ′ (𝐹) = 𝑘𝐻 × 𝑦(𝐻) .
S27
Relative Abundance
1.0
0.8
D
F11
A
B
F
C
G
E
H
0.6
0.4
0.2
0.0
0
2
4
6
8
10
12
14
16
Time (min)
Fig. S18. Relative abundance of different conformer types for the PPI→PPII transition in 10:88:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (F11) that does not work
for fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B, blue is conformer C, magenta is conformer D, green is conformer E, orange is conformer F, violet is
conformer G, and olive is conformer H. Fitting statistics are provided in Table S7, and the lines shown here represent
the best fitting results. The mathematical functions used to fit the data points were created by solving a series of
differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (A) = −𝑘𝐴 ×
𝑦(A) , 𝑦 ′ (𝐵) = 𝑘𝐴 × 𝑦(A) − 𝑘𝐵 × 𝑦(𝐵) , 𝑦 ′ (𝐶) = 𝑘𝐵 × 𝑦(𝐵) − 𝑘𝐶 × 𝑦(𝐶) , 𝑦 ′ (𝐷) = 𝑘𝐶𝐷 × 𝑦(𝐶) − 𝑘𝐷 × 𝑦(𝐷) , 𝑦 ′ (𝐸) = 𝑘𝐶𝐸 × 𝑦(𝐶) −
𝑘𝐸 × 𝑦(𝐸) , 𝑦 ′ (𝐺) = 𝑘𝐷 × 𝑦(𝐷) + 𝑘𝐸 × 𝑦(𝐸) − 𝑘𝐺 × 𝑦(𝐺) , 𝑦 ′ (𝐹) = 𝑘𝐺𝐹 × 𝑦(𝐺) , 𝑦 ′ (𝐻) = 𝑘𝐺𝐻 × 𝑦(𝐺) .
S28
Relative Abundance
1.0
F12
0.8
A
D
B
E
C
F
H
G
0.6
0.4
0.2
0.0
0
2
4
6
8
10
12
14
16
Time (min)
Fig. S19. Relative abundance of different conformer types for the PPI→PPII transition in 10:88:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (F12) that does not work
for fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B, blue is conformer C, magenta is conformer D, green is conformer E, orange is conformer F, violet is
conformer G, and olive is conformer H. Fitting statistics are provided in Table S7, and the lines shown here represent
the best fitting results. The mathematical functions used to fit the data points were created by solving a series of
differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (A) = −𝑘𝐴 ×
𝑦(A) , 𝑦 ′ (𝐵) = 𝑘𝐴 × 𝑦(A) − 𝑘𝐵 × 𝑦(𝐵) , 𝑦 ′ (𝐶) = 𝑘𝐵 × 𝑦(𝐵) − 𝑘𝐶 × 𝑦(𝐶) , 𝑦 ′ (𝐷) = 𝑘𝐶𝐷 × 𝑦(𝐶) − 𝑘𝐷 × 𝑦(𝐷) , 𝑦 ′ (𝐸) = 𝑘𝐶𝐸 × 𝑦(𝐶) −
𝑘𝐸 × 𝑦(𝐸) , 𝑦 ′ (𝐺) = 𝑘𝐶𝐺 × 𝑦(𝐶) − 𝑘𝐺 × 𝑦(𝐺) , 𝑦 ′ (𝐻) = 𝑘𝐷 × 𝑦(𝐷) + 𝑘𝐸 × 𝑦(𝐸) + 𝑘𝐺 × 𝑦(𝐺) − 𝑘𝐻 × 𝑦(𝐻) , 𝑦 ′ (𝐹) = 𝑘𝐻 × 𝑦(𝐻) .
S29
Relative Abundance
1.0
F13
A
0.8
F
B
C
D
E
G
H
0.6
0.4
0.2
0.0
0
2
4
6
8
10
12
14
16
Time (min)
Fig. S20. Relative abundance of different conformer types for the PPI→PPII transition in 10:88:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (F13) that does not work
for fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B, blue is conformer C, magenta is conformer D, green is conformer E, orange is conformer F, violet is
conformer G, and olive is conformer H. Fitting statistics are provided in Table S7, and the lines shown here represent
the best fitting results. The mathematical functions used to fit the data points were created by solving a series of
differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (A) = −𝑘𝐴 ×
𝑦(A) , 𝑦 ′ (𝐵) = 𝑘𝐴 × 𝑦(A) − 𝑘𝐵 × 𝑦(𝐵) , 𝑦 ′ (𝐶) = 𝑘𝐵 × 𝑦(𝐵) − 𝑘𝐶 × 𝑦(𝐶) , 𝑦 ′ (𝐷) = 𝑘𝐶 × 𝑦(𝐶) − 𝑘𝐷 × 𝑦(𝐷) , 𝑦 ′ (𝐸) = 𝑘𝐷 × 𝑦(𝐷) −
𝑘𝐸 × 𝑦(𝐸) , 𝑦 ′ (𝐺) = 𝑘𝐸 × 𝑦(𝐸) − 𝑘𝐺 × 𝑦(𝐺) , 𝑦 ′ (𝐹) = 𝑘𝐺𝐹 × 𝑦(𝐺) , 𝑦′ (𝐻) = 𝑘𝐺𝐻 × 𝑦(𝐺) .
S30
Relative Abundance
1.0
D*
R1
0.8
E*
F
A
C*
B*
G*
0.6
H*
0.4
0.2
0.0
0
30
60
90
120
150
180
210
Time (min)
Fig. S21. Relative abundance of different conformer types for the PPII→PPI transition in 97:1:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (R1) that works for
fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B*, blue is conformer C*, magenta is conformer D*, green is conformer E*, orange is conformer F, violet
is conformer G*, and olive is conformer H*. Fitting statistics are provided in Table S8, and the lines shown here
represent the best fitting results. The mathematical functions used to fit the data points were created by solving a series
of differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (F) =
−𝑘𝐹 × 𝑦(F) , 𝑦 ′ (𝐺∗) = 𝑘𝐹𝐺∗ × 𝑦(F) − 𝑘𝐺∗ × 𝑦(𝐺∗) , 𝑦 ′ (𝐻∗) = 𝑘𝐹𝐻∗ × 𝑦(F) − 𝑘𝐻∗ × 𝑦(𝐻∗) , 𝑦 ′ (𝐷∗) = 𝑘𝐹𝐷∗ × 𝑦(F) + 𝑘𝐺∗𝐷∗ × 𝑦(𝐺∗) +
𝑘𝐻∗𝐷∗ × 𝑦(𝐻∗) − 𝑘𝐷∗ × 𝑦(𝐷∗) , 𝑦 ′ (𝐸∗) = 𝑘𝐹𝐸∗ × 𝑦(F) + 𝑘𝐺∗𝐸∗ × 𝑦(𝐺∗) + 𝑘𝐻∗𝐸∗ × 𝑦(𝐻∗) − 𝑘𝐸∗ × 𝑦(𝐸∗) , 𝑦 ′ (𝐶∗) = 𝑘𝐷∗ × 𝑦(𝐷∗)
+ 𝑘𝐸∗ × 𝑦(𝐸∗) − 𝑘𝐶∗ × 𝑦(𝐶∗) , 𝑦 ′ (𝐵∗) = 𝑘𝐶∗𝐵∗ × 𝑦(𝐶∗) , 𝑦 ′ (𝐴) = 𝑘𝐶∗𝐴 × 𝑦(𝐶∗) .
S31
1.0
R2
D*
F
E*
Relative Abundance
A
0.8
C*
B*
G*
0.6
H*
0.4
0.2
0.0
0
30
60
90
120
150
180
210
Time (min)
Fig. S22. Relative abundance of different conformer types for the PPII→PPI transition in 97:1:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (R2) that works for
fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B*, blue is conformer C*, magenta is conformer D*, green is conformer E*, orange is conformer F, violet
is conformer G*, and olive is conformer H*. Fitting statistics are provided in Table S8, and the lines shown here
represent the best fitting results. The mathematical functions used to fit the data points were created by solving a series
of differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (F) =
−𝑘𝐹 × 𝑦(F) , 𝑦 ′ (𝐷∗) = 𝑘𝐹𝐷∗ × 𝑦(F) − 𝑘𝐷∗ × 𝑦(𝐷∗) , 𝑦 ′ (𝐸∗) = 𝑘𝐹𝐸∗ × 𝑦(F) − 𝑘𝐸∗ × 𝑦(𝐸∗) , 𝑦 ′
𝑦′
= 𝑘𝐹𝐻∗ × 𝑦(F) − 𝑘𝐻∗ × 𝑦(𝐻∗) ,
𝑘𝐶∗𝐵∗ × 𝑦(𝐶∗) , 𝑦 ′ (𝐴) = 𝑘𝐶∗𝐴 × 𝑦(𝐶∗) .
(𝐻∗)
𝑦′
(𝐶∗)
(𝐺∗)
= 𝑘𝐹𝐺∗ × 𝑦(F) − 𝑘𝐺∗ × 𝑦(𝐺∗) ,
= 𝑘𝐷∗ × 𝑦(𝐷∗) + 𝑘𝐸∗ × 𝑦(𝐸∗) + 𝑘𝐺∗ × 𝑦(𝐺∗) + 𝑘𝐻∗ × 𝑦(𝐻∗) − 𝑘𝐶∗ × 𝑦(𝐶∗) , 𝑦 ′ (𝐵∗) =
S32
1.0
R3
D*
A
Relative Abundance
C*
0.8
E*
F
B*
G*
0.6
H*
0.4
0.2
0.0
0
30
60
90
120
150
180
210
Time (min)
Fig. S23. Relative abundance of different conformer types for the PPII→PPI transition in 97:1:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (R3) that works for
fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B*, blue is conformer C*, magenta is conformer D*, green is conformer E*, orange is conformer F, violet
is conformer G*, and olive is conformer H*. Fitting statistics are provided in Table S8, and the lines shown here
represent the best fitting results. The mathematical functions used to fit the data points were created by solving a series
of differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (F) =
−𝑘𝐹 × 𝑦(F) , 𝑦 ′ (𝐺∗) = 𝑘𝐹𝐺∗ × 𝑦(F) − 𝑘𝐺∗ × 𝑦(𝐺∗) , 𝑦 ′ (𝐻∗) = 𝑘𝐹𝐻∗ × 𝑦(F) − 𝑘𝐻∗ × 𝑦(𝐻∗) , 𝑦 ′ (𝐷∗) = 𝑘𝐹𝐷∗ × 𝑦(F) + 𝑘𝐺∗ × 𝑦(𝐺∗) −
𝑘𝐷∗ × 𝑦(𝐷∗) , 𝑦 ′ (𝐸∗) = 𝑘𝐹𝐸∗ × 𝑦(F) + 𝑘𝐻∗ × 𝑦(𝐻∗) − 𝑘𝐸∗ × 𝑦(𝐸∗) , 𝑦 ′ (𝐶∗) = 𝑘𝐷∗ × 𝑦(𝐷∗) + 𝑘𝐸∗ × 𝑦(𝐸∗) − 𝑘𝐶∗ × 𝑦(𝐶∗) ,
𝑦 ′ (𝐵∗) = 𝑘𝐶∗𝐵∗ × 𝑦(𝐶∗) , 𝑦 ′ (𝐴) = 𝑘𝐶∗𝐴 × 𝑦(𝐶∗) .
S33
1.0
R4
D*
A
Relative Abundance
C*
0.8
E*
F
B*
G*
0.6
H*
0.4
0.2
0.0
0
30
60
90
120
150
180
210
Time (min)
Fig. S24. Relative abundance of different conformer types for the PPII→PPI transition in 97:1:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (R4) that works for
fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B*, blue is conformer C*, magenta is conformer D*, green is conformer E*, orange is conformer F, violet
is conformer G*, and olive is conformer H*. Fitting statistics are provided in Table S8, and the lines shown here
represent the best fitting results. The mathematical functions used to fit the data points were created by solving a series
of differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (F) =
−𝑘𝐹 × 𝑦(F) , 𝑦 ′ (𝐺∗) = 𝑘𝐹𝐺∗ × 𝑦(F) − 𝑘𝐺∗ × 𝑦(𝐺∗) , 𝑦 ′ (𝐻∗) = 𝑘𝐹𝐻∗ × 𝑦(F) − 𝑘𝐻∗ × 𝑦(𝐻∗) , 𝑦 ′ (𝐷∗) = 𝑘𝐹𝐷∗ × 𝑦(F) + 𝑘𝐻∗ × 𝑦(𝐻∗) −
𝑘𝐷∗ × 𝑦(𝐷∗) , 𝑦 ′ (𝐸∗) = 𝑘𝐹𝐸∗ × 𝑦(F) + 𝑘𝐺∗ × 𝑦(𝐺∗) − 𝑘𝐸∗ × 𝑦(𝐸∗) , 𝑦 ′ (𝐶∗) = 𝑘𝐷∗ × 𝑦(𝐷∗) + 𝑘𝐸∗ × 𝑦(𝐸∗) − 𝑘𝐶∗ × 𝑦(𝐶∗) ,
𝑦 ′ (𝐵∗) = 𝑘𝐶∗𝐵∗ × 𝑦(𝐶∗) , 𝑦 ′ (𝐴) = 𝑘𝐶∗𝐴 × 𝑦(𝐶∗) .
S34
Relative Abundance
1.0
R5
G*
F
0.8
A
E*
D*
C*
B*
H*
0.6
0.4
0.2
0.0
0
30
60
90
120
150
180
210
Time (min)
Fig. S25. Relative abundance of different conformer types for the PPII→PPI transition in 97:1:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (R5) that works for
fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B*, blue is conformer C*, magenta is conformer D*, green is conformer E*, orange is conformer F, violet
is conformer G*, and olive is conformer H*. Fitting statistics are provided in Table S8, and the lines shown here
represent the best fitting results. The mathematical functions used to fit the data points were created by solving a series
of differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (F) =
−𝑘𝐹 × 𝑦(F) , 𝑦 ′ (𝐺∗) = 𝑘𝐹𝐺∗ × 𝑦(F) − 𝑘𝐺∗ × 𝑦(𝐺∗) , 𝑦 ′ (𝐻∗) = 𝑘𝐹𝐻∗ × 𝑦(F) − 𝑘𝐻∗ × 𝑦(𝐻∗) , 𝑦 ′ (𝐸∗) = 𝑘𝐺∗ × 𝑦(𝐺∗) − 𝑘𝐸∗ × 𝑦(𝐸∗) ,
𝑦 ′ (𝐷∗) = 𝑘𝐸∗ × 𝑦(𝐸∗) + 𝑘𝐻∗ × 𝑦(𝐻∗) − 𝑘𝐷∗ × 𝑦(𝐷∗) , 𝑦 ′ (𝐶∗) = 𝑘𝐷∗ × 𝑦(𝐷∗) − 𝑘𝐶∗ × 𝑦(𝐶∗) , 𝑦 ′ (𝐵∗) = 𝑘𝐶∗𝐵∗ × 𝑦(𝐶∗) , 𝑦 ′ (𝐴) = 𝑘𝐶∗𝐴 ×
𝑦(𝐶∗) .
S35
Relative Abundance
1.0
R6
0.8
G*
F
A
E*
D*
C*
H*
B*
0.6
0.4
0.2
0.0
0
30
60
90
120
150
180
210
Time (min)
Fig. S26. Relative abundance of different conformer types for the PPII→PPI transition in 97:1:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (R6) that works for
fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B*, blue is conformer C*, magenta is conformer D*, green is conformer E*, orange is conformer F, violet
is conformer G*, and olive is conformer H*. Fitting statistics are provided in Table S8, and the lines shown here
represent the best fitting results. The mathematical functions used to fit the data points were created by solving a series
of differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (F) =
−𝑘𝐹 × 𝑦(F) , 𝑦 ′ (𝐺∗) = 𝑘𝐹𝐺∗ × 𝑦(F) − 𝑘𝐺∗ × 𝑦(𝐺∗) , 𝑦 ′ (𝐻∗) = 𝑘𝐹𝐻∗ × 𝑦(F) − 𝑘𝐻∗ × 𝑦(𝐻∗) , 𝑦 ′ (𝐸∗) = 𝑘𝐺∗ × 𝑦(𝐺∗) + 𝑘𝐻∗ × 𝑦(𝐻∗) −
𝑘𝐸∗ × 𝑦(𝐸∗) , 𝑦 ′ (𝐷∗) = 𝑘𝐸∗ × 𝑦(𝐸∗) − 𝑘𝐷∗ × 𝑦(𝐷∗) , 𝑦 ′ (𝐶∗) = 𝑘𝐷∗ × 𝑦(𝐷∗) − 𝑘𝐶∗ × 𝑦(𝐶∗) , 𝑦 ′ (𝐵∗) = 𝑘𝐶∗𝐵∗ × 𝑦(𝐶∗) , 𝑦 ′ (𝐴) = 𝑘𝐶∗𝐴 ×
𝑦(𝐶∗) .
S36
Relative Abundance
1.0
R7
0.8
G*
F
A
D*
E*
C*
H*
B*
0.6
0.4
0.2
0.0
0
30
60
90
120
150
180
210
Time (min)
Fig. S27. Relative abundance of different conformer types for the PPII→PPI transition in 97:1:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (R7) that does not work
for fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B*, blue is conformer C*, magenta is conformer D*, green is conformer E*, orange is conformer F, violet
is conformer G*, and olive is conformer H*. Fitting statistics are provided in Table S8, and the lines shown here
represent the best fitting results. The mathematical functions used to fit the data points were created by solving a series
of differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (F) =
−𝑘𝐹 × 𝑦(F) , 𝑦 ′ (𝐺∗) = 𝑘𝐹𝐺∗ × 𝑦(F) − 𝑘𝐺∗ × 𝑦(𝐺∗) , 𝑦 ′ (𝐻∗) = 𝑘𝐹𝐻∗ × 𝑦(F) − 𝑘𝐻∗ × 𝑦(𝐻∗) , 𝑦 ′ (𝐷∗) = 𝑘𝐺∗ × 𝑦(𝐺∗) + 𝑘𝐻∗ × 𝑦(𝐻∗) −
𝑘𝐷∗ × 𝑦(𝐷∗) , 𝑦 ′ (𝐸∗) = 𝑘𝐷∗ × 𝑦(𝐷∗) − 𝑘𝐸∗ × 𝑦(𝐸∗) , 𝑦 ′ (𝐶∗) = 𝑘𝐸∗ × 𝑦(𝐸∗) − 𝑘𝐶∗ × 𝑦(𝐶∗) , 𝑦 ′ (𝐵∗) = 𝑘𝐶∗𝐵∗ × 𝑦(𝐶∗) , 𝑦 ′ (𝐴) = 𝑘𝐶∗𝐴 ×
𝑦(𝐶∗) .
S37
Relative Abundance
1.0
R8
G*
A
E*
C*
F
0.8
H*
D*
B*
0.6
0.4
0.2
0.0
0
30
60
90
120
150
180
210
Time (min)
Fig. S28. Relative abundance of different conformer types for the PPII→PPI transition in 97:1:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (R8) that does not work
for fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B*, blue is conformer C*, magenta is conformer D*, green is conformer E*, orange is conformer F, violet
is conformer G*, and olive is conformer H*. Fitting statistics are provided in Table S8, and the lines shown here
represent the best fitting results. The mathematical functions used to fit the data points were created by solving a series
of differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (F) =
−𝑘𝐹 × 𝑦(F) , 𝑦 ′ (𝐺∗) = 𝑘𝐹𝐺∗ × 𝑦(F) − 𝑘𝐺∗ × 𝑦(𝐺∗) , 𝑦 ′ (𝐻∗) = 𝑘𝐹𝐻∗ × 𝑦(F) − 𝑘𝐻∗ × 𝑦(𝐻∗) , 𝑦 ′ (𝐸∗) = 𝑘𝐺∗ × 𝑦(𝐺∗) − 𝑘𝐸∗ ×
𝑦(𝐸∗) , 𝑦 ′ (𝐷∗) = 𝑘𝐻∗ × 𝑦(𝐻∗) − 𝑘𝐷∗ × 𝑦(𝐷∗) , 𝑦 ′ (𝐶∗) = 𝑘𝐷∗ × 𝑦(𝐷∗) + 𝑘𝐸∗ × 𝑦(𝐸∗) − 𝑘𝐶∗ × 𝑦(𝐶∗) , 𝑦 ′ (𝐵∗) = 𝑘𝐶∗𝐵∗ × 𝑦(𝐶∗) , 𝑦 ′ (𝐴) =
𝑘𝐶∗𝐴 × 𝑦(𝐶∗) .
S38
Relative Abundance
1.0
R9
0.8
A
D*
F
G*
C*
H*
B*
E*
0.6
0.4
0.2
0.0
0
30
60
90
120
150
180
210
Time (min)
Fig. S29. Relative abundance of different conformer types for the PPII→PPI transition in 97:1:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (R9) that does not work
for fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B*, blue is conformer C*, magenta is conformer D*, green is conformer E*, orange is conformer F, violet
is conformer G*, and olive is conformer H*. Fitting statistics are provided in Table S8, and the lines shown here
represent the best fitting results. The mathematical functions used to fit the data points were created by solving a series
of differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (F) =
−𝑘𝐹 × 𝑦(F) , 𝑦 ′ (𝐺∗) = 𝑘𝐹 × 𝑦(F) − 𝑘𝐺∗ × 𝑦(𝐺∗) , 𝑦 ′ (𝐻∗) = 𝑘𝐺∗ × 𝑦(𝐺∗) − 𝑘𝐻∗ × 𝑦(𝐻∗) , 𝑦 ′ (𝐸∗) = 𝑘𝐻∗𝐸∗ × 𝑦(𝐻∗) − 𝑘𝐸∗ ×
𝑦(𝐸∗) , 𝑦 ′ (𝐷∗) = 𝑘𝐻∗𝐷∗ × 𝑦(𝐻∗) − 𝑘𝐷∗ × 𝑦(𝐷∗) , 𝑦 ′ (𝐶∗) = 𝑘𝐷∗ × 𝑦(𝐷∗) + 𝑘𝐸∗ × 𝑦(𝐸∗) − 𝑘𝐶∗ × 𝑦(𝐶∗) , 𝑦 ′ (𝐵∗) = 𝑘𝐶∗𝐵∗ × 𝑦(𝐶∗) ,
𝑦 ′ (𝐴) = 𝑘𝐶∗𝐴 × 𝑦(𝐶∗) .
S39
Relative Abundance
1.0
R10
0.8
F
A
G*
H*
E*
D*
C*
B*
0.6
0.4
0.2
0.0
0
30
60
90
120
150
180
210
Time (min)
Fig. S30. Relative abundance of different conformer types for the PPII→PPI transition in 97:1:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (R10) that does not work
for fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B*, blue is conformer C*, magenta is conformer D*, green is conformer E*, orange is conformer F, violet
is conformer G*, and olive is conformer H*. Fitting statistics are provided in Table S8, and the lines shown here
represent the best fitting results. The mathematical functions used to fit the data points were created by solving a series
of differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (F) =
−𝑘𝐹 × 𝑦(F) , 𝑦 ′ (𝐺∗) = 𝑘𝐹 × 𝑦(F) − 𝑘𝐺∗ × 𝑦(𝐺∗) , 𝑦 ′ (𝐻∗) = 𝑘𝐺∗ × 𝑦(𝐺∗) − 𝑘𝐻∗ × 𝑦(𝐻∗) , 𝑦 ′ (𝐸∗) = 𝑘𝐻∗ × 𝑦(𝐻∗) − 𝑘𝐸∗ ×
𝑦(𝐸∗) , 𝑦 ′ (𝐷∗) = 𝑘𝐸∗ × 𝑦(𝐸∗) − 𝑘𝐷∗ × 𝑦(𝐷∗) , 𝑦 ′ (𝐶∗) = 𝑘𝐷∗ × 𝑦(𝐷∗) − 𝑘𝐶∗ × 𝑦(𝐶∗) , 𝑦 ′ (𝐵∗) = 𝑘𝐶∗𝐵∗ × 𝑦(𝐶∗) , 𝑦 ′ (𝐴) = 𝑘𝐶∗𝐴 × 𝑦(𝐶∗) .
S40
Relative Abundance
1.0
R11
F
0.8
G*
H*
E*
D*
C*
120
150
B*
A
0.6
0.4
0.2
0.0
0
30
60
90
180
210
Time (min)
Fig. S31. Relative abundance of different conformer types for the PPII→PPI transition in 97:1:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (R11) that does not work
for fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B*, blue is conformer C*, magenta is conformer D*, green is conformer E*, orange is conformer F, violet
is conformer G*, and olive is conformer H*. Fitting statistics are provided in Table S8, and the lines shown here
represent the best fitting results. The mathematical functions used to fit the data points were created by solving a series
of differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (F) =
−𝑘𝐹 × 𝑦(F) , 𝑦 ′ (𝐺∗) = 𝑘𝐹 × 𝑦(F) − 𝑘𝐺∗ × 𝑦(𝐺∗) , 𝑦 ′ (𝐻∗) = 𝑘𝐺∗ × 𝑦(𝐺∗) − 𝑘𝐻∗ × 𝑦(𝐻∗) , 𝑦 ′ (𝐸∗) = 𝑘𝐻∗ × 𝑦(𝐻∗) − 𝑘𝐸∗ × 𝑦(𝐸∗) ,
𝑦 ′ (𝐷∗) = 𝑘𝐸∗ × 𝑦(𝐸∗) − 𝑘𝐷∗ × 𝑦(𝐷∗) , 𝑦 ′ (𝐶∗) = 𝑘𝐷∗ × 𝑦(𝐷∗) − 𝑘𝐶∗ × 𝑦(𝐶∗) , 𝑦 ′ (𝐵∗) = 𝑘𝐶∗ × 𝑦(𝐶∗) − 𝑘𝐵∗ × 𝑦(𝐵∗) , 𝑦 ′ (𝐴) = 𝑘𝐵∗ ×
𝑦(𝐵∗) .
S41
Relative Abundance
1.0
R12
F
0.8
H*
G*
E*
D*
C*
120
150
B*
A
0.6
0.4
0.2
0.0
0
30
60
90
180
210
Time (min)
Fig. S32. Relative abundance of different conformer types for the PPII→PPI transition in 97:1:2 1propanol:H2O:HOAc (v:v:v) at 45 °C as a function of transition time and one transition route (R11) that does not work
for fitting the data points. Various colors are used to represent different conformations: black is conformer A, red is
conformer B*, blue is conformer C*, magenta is conformer D*, green is conformer E*, orange is conformer F, violet
is conformer G*, and olive is conformer H*. Fitting statistics are provided in Table S8, and the lines shown here
represent the best fitting results. The mathematical functions used to fit the data points were created by solving a series
of differential equations that describe the reaction rate of each conformer involved in the transition route: 𝑦 ′ (F) =
−𝑘𝐹 × 𝑦(F) , 𝑦 ′ (𝐻∗) = 𝑘𝐹 × 𝑦(F) − 𝑘𝐻∗ × 𝑦(𝐻∗) ,
𝑦 ′ (𝐺∗) = 𝑘𝐻∗ × 𝑦(𝐻∗) − 𝑘𝐺∗ × 𝑦(𝐺∗) , 𝑦 ′ (𝐸∗) = 𝑘𝐺∗ × 𝑦(𝐺∗) − 𝑘𝐸∗ ×
𝑦(𝐸∗) , 𝑦 ′ (𝐷∗) = 𝑘𝐸∗ × 𝑦(𝐸∗) − 𝑘𝐷∗ × 𝑦(𝐷∗) , 𝑦 ′ (𝐶∗) = 𝑘𝐷∗ × 𝑦(𝐷∗) − 𝑘𝐶∗ × 𝑦(𝐶∗) , 𝑦 ′ (𝐵∗) = 𝑘𝐶∗ × 𝑦(𝐶∗) − 𝑘𝐵∗ × 𝑦(𝐵∗) , 𝑦 ′ (𝐴) =
𝑘𝐵∗ × 𝑦(𝐵∗) .
S42
−6
(𝑘) =
−𝐸𝑎
+
( )
−8
−10
−12
−14
−16
3.1
F
F
G*
H*
E*
C*
E*
H*
E*
E*
C*
B*
3.2
3.3
3.4
1/T
103
3.5
3.6
(K−1)
Fig. S33. Arrhenius plot of the rate constants for six transition steps involved in the PPII→PPI transition of Pro13 at
five different temperatures (5, 15, 23, 35, and 45 °C). Error bars represent the standard deviation from quadruplicate
analysis.
S43
Normalized Intensity
H
G F E D
C B
A
45 C
1500
42.5 C
1600
40 C
400
37.5 C
300
35 C
1500
32.5 C
1500
30 C
400
15 C
700
250
270
290
310
330
350
CCS (Å2)
Fig. S34. Collision cross section distributions for the [Pro13+2H]2+ ions obtained at equilibrium of the PPI→PPII
transition over a temperature range of 15–45 οC. The low-abundance conformers present within the equilibriums
distribution are shown as red insets. The transition temperatures for the obtained distributions are indicated in each
trace, and dashed lines delineate the collision cross section region for each conformer type.
S44
Normalized Intensity
H* G* F E*D* C* B*
250
A
100
45 C
40
42.5 C
40
40 C
40
37.5 C
60
35 C
50
32.5 C
110
30 C
50
15 C
270
290
310
330
350
CCS (Å2)
Fig. S35. Collision cross section distributions for the [Pro13+2H]2+ ions obtained at equilibrium of the PPII→PPI
transition over a temperature range of 15–45 οC. The low-abundance conformers present within the equilibriums
distribution are shown as red insets. The transition temperatures for the obtained distributions are indicated in each
trace, and dashed lines delineate the collision cross section region for each conformer type.
S45
Reference:
1. Shi, L., Holliday, A.E., Shi, H., Zhu, F., Ewing, M.A., Russell, D.H., Clemmer, D.E.:
Characterizing intermediates along the transition from polyproline I to polyproline II using ion
mobility spectrometry-mass spectrometry. J. Am. Chem. Soc. 136, 12702–12711 (2014)
S46