30. 10. 2017

Utopia

W. Bermel

6.-9. November 2017

Deutschland

relaxation delay

1 - 3 s

seq (pulsesdelays...)

10 - 300ms

acq

100ms

NMR experiment

A. Viegas, T. Viennet, T.Y. Yu, F. Schumann, W. Bermel,

G. Wagner & M., Etzkorn,J. Biomol. NMR 64, 9-15 (2016)

UTOPIA - NMRunified time-optimized interleaved acquisition NMR

Utopia

3

relaxation delay

1 - 3 s

seq (pulsesdelays...)

10 - 300ms

acq

100ms

NMR experiment

Utopia

4

rel. delayseq 1 acq 1

UTOPIA experiment

seq 2 acq 2 .......

Utopia

5

rel. delay1 1

UTOPIA experiment

NUC(e) NUC(d) .......

- orthogonal

- unterschiedliche NUC(e)

- (unterschiedlicheNUC(d) )

- vergeichbare Empfindlichkeit

- komplementäre Information

NUC(e)

2 2

NUC(d)

H

NN

H

H

Cab

N

CO

(H)CbCa(CON)H HNCON

H H

Cb - Hnb

| | |

|

Ha

N- CO - N - Ca - CO - -

t1

t2 t1

t1

COCO

t2

t2

Utopia

7

H

Cab

N

CO

(H)CbCa(CON)H HNCON

CON vor (H)CbCa(CON)H(H)CbCa(CON)H pulse auf 13C, (15N) =>

HN vor (H)CbCa(CON)H,

HN 1H Entkopplung => HN als TROSY

13C Entkopplung CON vor HN

CON 1H Entkopplung

=>pulse auf 1H

=>

=> 180° 1H Pulse in t1

mit HN selektiven Pulsen

Utopia

8

H

Cab

N

CO

(H)CbCa(CON)HHNCON

best-HN =>

CON =>180° 1H Pulse in t1

HN selektive Pulse

Effekt auf HN

(1H Entkopplung) => Effekt auf HN

Effekt auf (H)CbCa(CON)H

Effekt auf CON

Utopia

9

¾UV¨©©©©©©©©©©©©©©ª

ÊU$l'l$VV<UUV$l'lUM.

ÀUUU'UU$c'•$UU'UV¨¬ª

ÅUUUUUV²UUUV²UUUUUUV

¾UUUUUUU)VW)UUUUUUUU

ÊU$l'l$VV<UUV$l'lUM.

ÀUUU'UU$c'•$UU'UV¨¬ª

ÅUUUUUV²UUUV²UUUUUUV

CON

dec.

dec.

dec.

BIP

Utopia

10

167169170171172173174175176177178179[ppm] 168

ca. 80%

CON

1H decoupling

BIP pulses

CON projection

Utopia

11

HSQC test

rel. delaytest seq 100ms HSQC acq

hsqcetgpsp

Utopia

12

8 7 6 5 4 3 2 1 0[ppm]

crp20,1.5,20.1

reference

BIP - 3.4ms - BIP - 50ms - AQT (100ms)AQT ((Crp)(p5m4)n)

BIP

decoupling 15-50%

98%

CON

Utopia

13

8 7 6 5 4 3 2 1 0[ppm]

reference

Eburp (4.4 ppm) + Reburp (4.4 ppm)

best-HN TROSY

@ 8.3ppm

Eburp (4.4 ppm) + Reburp (6.274 ppm)

Utopia

14

4.24.34.44.54.64.74.84.95.05.15.25.35.45.55.65.75.8 4.1[ppm]

reference

Eburp (4.4 ppm) + Reburp (4.4 ppm) ca. 70% (Ha)

best-HN TROSY

@ 8.3ppm

Eburp (4.4 ppm) + Reburp (6.274 ppm) ca. 40% (Ha)

Utopia

15

-2-112 11 10 9 8 7 6 5 4 3 2 1 0[ppm] -3

Reburp (4.4 ppm)

Reburp (6.274 ppm)

Eburp (4.4 ppm)

1D (zgesgp)

best-HN TROSY

Utopia

16

104106108110112114116118120122124126128[ppm] 102

best-HN TROSY (relaxation delay)

HN TOSY projection

D1

1.625s

0.4s ca. 80%

Utopia

17

(H)CbCa(CON)Hreference

Utopia ca 80%

6.06.57.07.58.08.59.09.510.0[ppm]

Utopia

18

167169170171172173174175176177178179[ppm] 168

reference (1H decoupling)

CON

Utopia ca. 75-80%

reference (BIP) ca. 75-80%

Utopia

19

104106108110112114116118120122124126128[ppm] 102

reference

Utopia ca 85-90%

best-HN TROSY

Utopia

20

Experiment-Zeit

(H)CbCa(CON)H

best-HN TROSY I (d1=1.625s)

CON

151 min

149 min

179 min

(ut 85%) 109 min

84 min

115 min

(ut 75%)

(ut 80%)

best-HN TROSY II (d1=0.4s)

*korrigiert für 80%

86 min * (ut 85%) 62 min

(H)CbCa(CON)H + CON + best-HN TROSY I

(H)CbCa(CON)H + CON + best-HN TROSY II

(H)CbCa(CON)H + CON

Utopia

308 min

261 min

199 min

153 min

Utopia

21

[ppm]

15

20

25

30

35

40

45

50

55

60

65

70

10

6.06.57.07.58.08.59.09.510.0[ppm]

[ppm]

106

110

114

118

122

126

102

168170172174176178[ppm]

(H)CbCa(CON)HUtopia

CON best-HN TROSY (transposed *)

* sr changed by 45Hz in both dimensions

Utopia

22

nus3D-(H)CbCa(CO)NH

Utopia

CON

best-HN TROSY

12.5% 128 x 64 x 2k

[ppm]

106

110

114

118

122

126

102

168170172174176178[ppm]

6.0

6.5

7.0

7.5

8.0

8.5

9.0

9.5

10.0

[ppm]

105110115120125[ppm]

F3 [ppm]9 8 7

F2

[pp

m]

14

01

30

12

01

10

F1

[pp

m]

80

60

Utopia

23

- CO - N - Ca - CO - -NNN

HH

H

Ca

N

CO

Cb - Hnb

| | |

|

Ha

COCO

3D-tr-HNCa

t1

t3

t2

A. Viegas, T. Viennet, T.Y. Yu, F. Schumann, W. Bermel, G. Wagner & M.,

Etzkorn,J. Biomol. NMR 64, 9-15 (2016)

CON

t1

t2

dec

Utopia

24

tr-HNCa

Utopia

CON

64 x 64 x 2k

F3 [ppm]9 8 7

F2

[pp

m]

14

01

30

12

01

10

F1

[pp

m]

65

60

55

[ppm]

168170172174176178[ppm]

104

106

108

110

112

114

116

118

120

122

124

126

128

130

102

64 (after Ipap) x 2k

Utopia

25

N- CO - N - Ca - CO - -

HH

H

Cali

N

CO

HCCH-TOCSY

Cb - Hnb

| | |

|

Ha

best-HNCO

t1

t3

t2

t2

t3t1

Utopia

26(single receiver, nbl=2 (st))

nus HCCH-TOCSY

reference Utopia

25% 128 x 128 (single receiver, nbl=2 (st); same nuslist, same filesize)

ca. 70-85% Haca. 96% sidechains

F3 [ppm]6 5 4 3 2 1

F2

[pp

m]

80

60

40

20

F1

[pp

m]

8

6

4

F3 [ppm]6 5 4 3 2 1

F2

[pp

m]

80

60

40

20

F1

[pp

m]

8

6

4

Utopia

27

nus best-HNCO

reference Utopia

ca. 85%

25% 128 x 128 (single receiver, nbl=2 (st); same nuslist, same filesize)

F3 [ppm]9 8 7

F2

[pp

m]

13

01

20

11

0

F1

[pp

m]

50

46

42

F3 [ppm]9 8 7

F2

[pp

m]

13

01

20

11

0

F1

[pp

m]

17

81

74

Utopia

28

CON + best-HN TROSY + 2D-(H)CbCa(CON)H

best-HNCO + HCCH-TOCSY

CON + best-HN TROSY + 3D-(H)CbCa(CO)NH

*A. Viegas, T. Viennet, T.Y. Yu, F. Schumann, W. Bermel, G. Wagner & M., Etzkorn,J. Biomol. NMR 64, 9-15 (2016)

3D-tr-HNCa + CON *

3D-15N NOESY-TROSY + CC-FLOPSY *

HH-COSY + 1D 13C *

HH-TOCSY + 1D 13C *

Utopia

29

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