the figures in this presentation were taken from: wadsworth
DESCRIPTION
The figures in this presentation were taken from: http://www.wadsworth.org/ - PowerPoint PPT PresentationTRANSCRIPT
![Page 1: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/1.jpg)
The figures in this presentation were taken from:http://www.wadsworth.org/Blachly-Dyson, E., Peng, S. Z., Colombini, M. and Forte, M. 1989. Probing the structure of the mitochondrial channel, VDAC, by site-directed mutagenesis: a progress report. Journal of Bioenergetics and Biomembranes, 21:471-483.Blachly-Dyson, E., Peng, S. Z., Colombini, M. and Forte, M. 1990. Alteration of the selectivity of the VDAC ion channel by site-directed mutagenesis: implications for the structure of a membrane ion channel. Science, 247: 1233-1236.Thomas, L., Kocsis, E., Colombini, M., Erbe, E., Trus, B.L. and Steven, A.C. 1991. Surface topography and molecular stoichiometry of the mitochondrial channel, VDAC, in crystalline arrays. Journal of Structural Biology, 106: 161-171.Peng, S., Blachly-Dyson, E., Colombini, M. and Forte, M. 1992. Large scale rearrangement of protein domains is associated with voltage gating of the VDAC channel. Biophysical Journal, 62: 123-153.Thomas, L., Blachly-Dyson, E., Colombini, M., and Forte, M. 1993. Mapping of residues forming the voltage sensor of the VDAC ion channel. Proceedings of the National Academy of Sciences U.S.A., 90: 5446-5449.Song, J., Midson, C., Blachly-Dyson, E., Forte, M., and Colombini, M. 1998. The sensor regions of VDAC are translocated from within the membrane to the surface during the gating processes. Biophysical Journal, 74: 2926-2944.Song, J., Midson, C., Blachly-Dyson, E., Forte, M., and Colombini, M. 1998. The topology of VDAC as probed by biotin modification. Journal of Biological Chemistry, 273:24406‑24413.Vander Heiden, M.G., Chandel, N.S., Li, X.X., Schumacker, P.T., Colombini, M., and Thompson, C.B. 2000. Outer mitochondrial membrane permeability can regulate coupled respiration and cell survival. Proceedings of the National Academy of Sciences U.S.A., 97: 4666‑4671.
![Page 2: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/2.jpg)
Mitochondria and Cellular Metabolism
![Page 3: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/3.jpg)
Voltage Dependent Anion-selective Channel (VDAC):
• 32kDa protein located in the mitochondrial outer membrane • Responsible for the metabolite permeation through the mitochondrial outer membrane of all eukaryotes
• Permeable to non-electrolytes up to 5 kDa but far more restrictive for charged molecules
![Page 4: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/4.jpg)
from: http://www.wadsworth.org/BMS/SCBlinks/channels2.htm
http://www.wadsworth.org/rvbc/tomography.html
work of Carmen Mannella
a mitochondrion...
![Page 5: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/5.jpg)
Biophysics of Channels Associated with Apoptosis
Images of VDAC
2D-crystals in mitochon-drial outer membrane
averaged surface view
Frozen-hydrated specimen
by Carmen Mannella
from: http://www.wadsworth.org
/BMS/SCBlinks/vdac.htm
![Page 6: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/6.jpg)
![Page 7: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/7.jpg)
human VDAC1
N. crassa VDAC
A V P
AF
SD
IA
KS A
ND
LL
NK
DF
YH
L
AA
G
T
I
E
V
K
S
N
T
P
N
N37
V
A
F
K
V
T
G
K
S
T
H
DK
V
T
S
G
A
L
E
G
K
F
T
D
K
P
N
G
L T68
V
T
Q
T
W
N
T
A
N
A
L
E
T
K
V
E
M
AD
N
L
A
K
G
L
K
A
E
G
I
F
S
F
L
P
A
T
N
A
R
GA
K111FN
L
H
F
K
Q
S
N
F
H
G
R
A
F
F
D
L
L
KG
P
T
A
N
I
D
A
I
V
G
H E144
G
F
L
A
G
A
S
A
G
Y
D
V
QKAAI
T
G Y S A A V
G
Y
H
A
P
T
Y
S
A
A
I
T
A
T
D
N
L
S
V
F
S189
AS Y Y H K V N S Q V E A G S K A
T
WN
SKTGNTVGLEVATK
Y
R
I
D
P
V
S
F
V
K
G
K
I
N
D
R
G
V
A
A
I
A
Y
N
V
L
LR
E252GV
T
L
G
V
G
A
S
F
D
T
Q
K
LD
Q
A
T
H
K
V
G
T
S
F
T
F
E
S282
A V P PTYA
D LGKS AR
D VFTK GYG F
G L IKLDLKTKSEN
GLEFTSSGSANTE
T TKVTGSLETKYRWTEYGLT
F TEKWNTDNTLGTEITVEDQ
LARGLKLTFDSSFSPNTGKKN
AKIKTGYKREHINLGCDMDFD
I AGPSIRGALVLGY E
GWLAGYQMNFETAKSRV
TQ S N F A V
GYKTDEFQLHTNVNDGTEFGGS I Y Q K V N K K L E T A V N L
AW
TAGNSNTRFGIAAKYQIDPDACFSAKVNN
SSLIGLGYTQTL
KPGIKLTLSALLDGKN
VNAGGHKLGLGLEFQA
![Page 8: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/8.jpg)
The functional properties of VDAC were analyzed by reconstituting the channels into a planar phospholipid membrane
• Detergent-solubilized VDAC is dispersed into the aqueous phase
• VDAC spontaneously inserts into the membrane
![Page 9: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/9.jpg)
Top view
Side view
Cross section
Saran partition0.1 mm hole
Chamber for making planar membranes
![Page 10: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/10.jpg)
0.1 mm
Formation of Membrane from Monolayers
![Page 11: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/11.jpg)
V
+
-
syringe
stir bar
electrode
cis trans
0.1 mm
Planar Membrane Setup
![Page 12: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/12.jpg)
200 pA
switch from 10 to 40 mV
return to 10 mV
addition of VDAC
time (sec)
Insertion and voltage-dependent closure of VDAC
![Page 13: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/13.jpg)
![Page 14: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/14.jpg)
The voltage dependence of VDAC
![Page 15: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/15.jpg)
![Page 16: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/16.jpg)
Time (minutes) 50 100 150 200 250
![Page 17: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/17.jpg)
Ion F
lux
(x105 io
ns/
sec )
0
1
2
3
4
5
6
7
Ion F
lux
( 106 io
ns/
sec )
0
2
4
6
8
10
12
14
16
VDAC openVDAC closed
Na+ PC succinate2- ATP4--2HPO4
VDAC Selectivity Change Upon Closure
2-
![Page 18: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/18.jpg)
current recording
applied voltage
![Page 19: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/19.jpg)
![Page 20: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/20.jpg)
![Page 21: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/21.jpg)
![Page 22: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/22.jpg)
Solution structure of the integral human membrane protein VDAC-1 in detergent micelles; Hiller et al., Science. 321: 1206–1210 (2008).
![Page 23: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/23.jpg)
Point mutations made in disputed strands
![Page 24: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/24.jpg)
![Page 25: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/25.jpg)
![Page 26: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/26.jpg)
![Page 27: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/27.jpg)
![Page 28: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/28.jpg)
![Page 29: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/29.jpg)
![Page 30: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/30.jpg)
![Page 31: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/31.jpg)
![Page 32: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/32.jpg)
![Page 33: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/33.jpg)
(25 mM)
![Page 34: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/34.jpg)
![Page 35: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/35.jpg)
![Page 36: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/36.jpg)
![Page 37: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/37.jpg)
![Page 38: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/38.jpg)
Biotin modification + Avidin binding
biotin
![Page 39: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/39.jpg)
![Page 40: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/40.jpg)
![Page 41: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/41.jpg)
![Page 42: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/42.jpg)
![Page 43: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/43.jpg)
![Page 44: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/44.jpg)
![Page 45: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/45.jpg)
![Page 46: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/46.jpg)
![Page 47: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/47.jpg)
![Page 48: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/48.jpg)
![Page 49: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/49.jpg)
![Page 50: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/50.jpg)
![Page 51: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/51.jpg)
![Page 52: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/52.jpg)
![Page 53: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/53.jpg)
![Page 54: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/54.jpg)
![Page 55: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/55.jpg)
![Page 56: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/56.jpg)
![Page 57: The figures in this presentation were taken from: wadsworth](https://reader035.vdocuments.us/reader035/viewer/2022062309/56813660550346895d9ded3e/html5/thumbnails/57.jpg)