riding the lipid tides
DESCRIPTION
Riding the lipid tides. David Stopar Wageningen, March-May 2003. CAUTION!!!. THE RESULTS AND MODELS PRESENTED HERE MAY BE PREMATURED AND UNSUFICIENTLY ANALISED. DIGESTION OF THESE RESULTS MAY LEAD TO SYSTEMIC EFFECTS. PRECAUTION : Avoid ingestion and digestion of this presentation. - PowerPoint PPT PresentationTRANSCRIPT
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Riding the lipid tides
David Stopar
Wageningen, March-May 2003
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CAUTION!!!
THE RESULTS AND MODELS PRESENTED HERE MAY BE
PREMATURED AND UNSUFICIENTLY ANALISED.
DIGESTION OF THESE RESULTS MAY LEAD TO SYSTEMIC
EFFECTS.
PRECAUTION: Avoid ingestion and digestion of this presentation
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PC18 1; 9-cis-octadecenoic
PC14 1; 9-cis-tetradecenoic
PC16 1; 9-cis-hexadecenoic
PC20 1; 11-cis-eicosenoic
PC22 1; 13-cis-docosenoic
Different PC’s
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Different mutants in DOPC
G3C A49C
T19C T46C
V29C A25C
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Major coat protein structural domains
1-5 N-terminus
6-16 amphiphatic helix
17-23 loop
24-46 transmembrane helix
47-50 C-terminus
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Different mutants in PC14 and PC22
G3CS13CA10C
Y21CV29C
V30C
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Different mutants in PC14 and PC22
65
60
55
50
2Azz
(G
)
2422201816141210
amino acid residue
60
55
50
45
40
35
2A
zz (
G)
40302010
amino acid residue
65
60
55
50
2Azz
(G
)
424038363432302826
amino acid residue
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CD spectra of major coat protein in different PC’c
6
4
2
0
-2
-4
arbi
trar
y un
its
250240230220210200
nm
pc14 pc16 pc18 pc20 pc22
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I39C mutant in a binary mixture of PC14 and
PC22
33803360334033203300
G
PC22
PC14
20/80
40/60
60/40
80/20
100/0
0/100
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Spin labeled T46C in different PC14/PC18 mixtures
64
63
62
61
2A
zz (
ga
uss
)
100806040200mol fraction of PC14
275 K
58.5
58.0
57.5
57.0
56.5
2A
zz (
ga
uss
)
100806040200mol fraction of PC14
295 K
55.5
55.0
54.5
54.0
53.5
53.0
52.5
2A
zz (
ga
uss
)
100806040200mol fraction PC 14
300 K30x10
-3
25
20
15
10
5
k (e
xp)
300295290285280275
temperature
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Phase transitions of different PC’s
66
64
62
60
58
56
54
52
G
310305300295290285280275
T (K)
PC22
PC14
-40
-30
-20
-10
0
10
20
Tm
22201816
PC
ns NaCl CaCl
16.2
16.0
15.8
15.6
Tm
10080604020
L/P
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Trp26 and BADAN T46C fluorescence in different PC’s
1.0
0.8
0.6
0.4
0.2
arbi
trar
y un
its
440420400380360340320300
max
1.030
1.025
1.020
1.015
1.010
1.005
1.000
max
PC14
/m
axPC
2220181614
PC
'trp14/trp' 'bad14/bad'
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Quenching studies
1.4
1.3
1.2
1.1
1.0
Fo/
F
14121086420
concentration of spin label
Trp quenching by spin label 1.4
1.3
1.2
1.1
1.0
Fo/
F14121086420
concentration of spin label
BADAN
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T46C reconstituted in different PC’s
63.0
62.5
62.0
61.5
61.0
60.5
60.0
2A
zz
2220181614
PC
arb
itra
ry u
nits
33803360334033203300[G]
PC14
PC16
PC18
PC20
PC22
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A49C reconstituted in PC14 and PC22
200x10-6
100
0
-100
-200
arbi
trar
y un
its
33803360334033203300
G
pc14 pc22
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Major coat protein in UREA, Cholate, SDS and TFE
-40
-20
0
20
arbi
trar
y un
its
250240230220210200nm
UREA
SDS
cho
TFE
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Different mutants in different solvents
Cholate
A25C
T46C
A7C
TFE SDS
DMSO
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That’s all we had time for, folks!
Thanks very much for your help and
cooperativity.
Cheers,
David