Upload
eara
View
50
Download
0
Embed Size (px)
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 Presentation
Citation preview
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
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
Different mutants in DOPC
G3C A49C
T19C T46C
V29C A25C
Major coat protein structural domains
1-5 N-terminus
6-16 amphiphatic helix
17-23 loop
24-46 transmembrane helix
47-50 C-terminus
Different mutants in PC14 and PC22
G3CS13CA10C
Y21CV29C
V30C
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
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
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
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
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
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'
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
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
A49C reconstituted in PC14 and PC22
200x10-6
100
0
-100
-200
arbi
trar
y un
its
33803360334033203300
G
pc14 pc22
Major coat protein in UREA, Cholate, SDS and TFE
-40
-20
0
20
arbi
trar
y un
its
250240230220210200nm
UREA
SDS
cho
TFE
Different mutants in different solvents
Cholate
A25C
T46C
A7C
TFE SDS
DMSO
That’s all we had time for, folks!
Thanks very much for your help and
cooperativity.
Cheers,
David