1. A mechanism for coupled gain of structure and loss of function in amyotrophic lateral sclerosis
Dr. Eamonn F. Healy
Professor of Chemistry
St. Edward’s University
Austin, Tx.

2. Structure and Function in SOD1-related Amyotrophic Lateral Sclerosis
Proof of Concept : desolvated h-bonds
Gain of interaction (GOI) in SOD1
Oxidation-induced gain of structure in SOD1

3. Tyrosine kinase inhibition: Ligand binding and conformational change in c-Kit and c-Abl
N-lobe
C-lobe

4. Tyrosine kinase inhibition: Ligand binding and conformational change in c-Kit and c-Abl
FEBS Lett , 583,

5. Acetylenic Inhibitors of ADAM10 and ADAM17: In silico analysis of potency and selectivity
loop
J. Mol. Graph. Model, 2010, 29,

6. The effect of desolvation on nucleophilic halogenase activity
Computational and Theoretical Chemistry, 2011, 964,

7. r=14 Wrapping a backbone hydrogen bond CHn (n= 1,2,3) backbone
Carbonyl O
Amide N
Amide H
a-carbon HB r
Extent of wrapping: r
r=14
desolvation
spheres

8. Fernandez, A. et al. PNAS 2003;100:
How many residues (n)
wrap a hydrogen bond?
desolvation sphere radius: r=6Å 1
periphery centrality pathogeny
ligand carriers complexes / signaling toxins / prions
enzymes modular hubs aggregates
<n> 2 1 2 1
Fernandez, A. et al. PNAS 2003;100:
& 2003;100: 2

9. A Model for heat shock inhibition of fibril formation in ataxin-3
Complex (PDB code) Y
d (10-3 Å-2)
Yint
dint (10-3 Å-2)
Ataxin-3/aB-crystallin (this work) 28
1.39 7
5.20
HLA antigen A-2/b2-microglobulin (1i4f) 36
1.58 6
6.50
Ig-light chain dimer (1jvk) 26
1.78 8
6.33
sHsp complex
aB-crystallin
ataxin-3
Cell Biochem. Biophys. 2014, 69,

10. Structure and Function in SOD1-related Amyotrophic Lateral Sclerosis
Proof of Concept : desolvated h-bonds
Gain of interaction (GOI) in SOD1
Oxidation-induced gain of structure in SOD1

11. Prudencio, M. Et et al. Hum. Mol. Gen. 2010;19:4774-4789
The aggregation of wt and fALS mutant SOD1
Prudencio, M. Et et al. Hum. Mol. Gen. 2010;19:

12. A Model for non-obligate oligomer formation in protein aggregation
SOD1 69
143
Biochem. Biophys. Res. Commun., 2015, 465 ,

13. The effect of superoxide dismutase (SOD1) electrostatic
loop dynamics on amyloid-like filament formation
Pathological
Physiological
Euro. Biophys. J., 2016, 45,

14. Biochem. Biophys. Res. Commun., 2016, 478, 1634-1639.
A mechanism for propagated SOD1 misfolding from frustration analysis of a G85R mutant protein assembly
Biochem. Biophys. Res. Commun., 2016, 478,

15. Frustration and enervation in SOD1
EL (A)
Cys57-Cys146
(A + F)
oligomer
RMSF color
(in Fig 4)
unconstrained
CysS SCys
apo wtSOD1 obligate homodimer
black
constrained
apo wtSOD1/mutant SOD1 non-obligate tetramer
cyan
Cys(SH)(HS)Cys
apo+reduced wtSOD1/mutant SOD1 non-obligate tetramer
green EL S
ZBL S
ZBL
Chain A S
Chain F S EL
Plos ONE , 2017, 12 (5), e

16. In silico modeling of solvated near-native conformers as an aid to understanding the propagated misfolding of SOD1
Chain A: zinc binding loop
Chain F: zinc binding loop
RMSF(Å)
RMSF(Å)
Chain A: electrostatic loop
Chain F: electrostatic loop
RMSF(Å)
RMSF(Å)

17. 1.1 ns 1.1 ns 1.4 ns wt 2 ns wt 2 ns 1.2 ns S134 S134 D83 D124 D125
P74
G138 W W
H80
T137
H80
E77 W W W
G127
D76
G127
R79 W W W
G72
T135 W W W
T135
G72
E78
K70 W
H71
K70 wt
2 ns
G138
D124
L126
D125
N139
T137
D125
G138
N139
T137 W
G127
K136
K136
L126
K128 W
1.2 ns W
T135
S134
D124
E133
G129
P74
G73
S134
K128 W W
1.1 ns
H71 W W
E132 W
G129 W
G138 W
E132
G130
N131
D76 W W W W
N131
G130
K128
H80 W W
K136
T135
G129

18. A prion mechanism for propagated SOD1 misfolding

19. Structure and Function in SOD1-related Amyotrophic Lateral Sclerosis
Proof of Concept : desolvated h-bonds
Gain of interaction (GOI) in SOD1
Oxidation-induced gain of structure in SOD1

20. A model for coupled gain of structure and loss of function in amyotrophic lateral sclerosis
RRM2 190
261
TDP-43

21. Pelmenschikov & Siegbahn Inorganic chemistry, 2005; 44, 3311-3320
SOD1 Dismutase cycle
Cu-His63=2.55 H
DG=2.4
TS 2 H
DG=-11.5 H O O O- O O. O
DG#=17.6
DG#=-11.5 H
+ O2-.
+ H+
Cu II
- H2O2
His deprotonation
Cu II
Cu oxidation
Cu I
-His61
H-His61
H-His61
Cu-Ne(His61)=2.55
TS 1
DG=7.8
DG#=12.6
DG=1.0
DG=0 -O O.
DG=0
HO.
H0-
Cu II
+ O2-.
+ H+
. O O
-O2
Cu I
+ H2O2
-His61
Cu II H
Cu II
H-His61
Cu II
-His61
-His61
DG#=31.4
H-His61
Cu-Zn=6.1
Cu-His61=2.2
Cu-Ne(His61)=2.31
Cu-Zn=6.6
Cu-His61=3.1
- O2-./ H+
+ H2O2
SOD1 Peroxidase cycle
Pelmenschikov & Siegbahn Inorganic chemistry, 2005; 44,

22. End on binding of hydrogen peroxide
His61
His46
His118
His44

23. Fenton Chemistry H
CuI-O
CuI-O O H O

24. -His61 -His61 - -His61 -His61 SOD1 Peroxidase cycle Cu II Cu I Cu I
LUMO
LUMO
CO3-.
H2O
Cu II
-His61
Homolytic
Cleavage
HOMO
HOMO
DE=-13.7
DG=+1.6
CO2- H O O
CO2-
H-His61 H H
-0.47 H H
-0.38 H
-0.20
Cu I O O
-0.29 O O
-0.27 O O
-0.36
H-His61
H-His61
H-His61
HO.
Cu I
Cu I
Cu I
H2O
+ H+
+CO2
Cu II
-His61
Heterolytic
Cleavage
DE=-5.6
DG=+7 H H
DE=-19.8
DG=-11.5 H - H H
DE=-0.9
DG=+11.8
DE=-11.9
DG=-2.5 O O O- O O
- H+
DE=-7.8
DG=-4.9 O O
+ H2O2 O
H-His61
H-His61 H
Cu I
Cu I
Cu II
Cu II
Cu II
-His61
-His61
H-His61 H H
DE=-1.4
DG=+11.3 O O
H-His61
Cu I
SOD1 Peroxidase cycle
+ HCO4- H
+ H2O2 O O.
DG=0
Cu I
Cu I
- O2-.
- H+
H-His61
H-His61

25. H69
H44
H69
K134
D122
N84
D122

26. Collaborators: Dr. Peter King ;
Acknowledgements
Collaborators: Dr. Peter King ;
Dr. Ariel Fernandez (Rice U)
Dr. Neil Cashman (U British Columbia)
Dr. Kevin Dalby (UT Austin)
Students: Jonathan Sanders; Skylar Johnson; Alicia Jones; Moises Mejia; Mina Nakhla;
Pablo Romano; Carley Little; Cassandra Petty;
Tony Velez; Luis Cervantes; Analise Roth-Rodriguez
Funding: National Institute of General Medical Services (1K12GM102745)
National Science Foundation (DUE )
The Welch Foundation (BH-0018)
W. M. Keck Foundation