Riding the lipid tides

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