This document discusses techniques for optimizing NMR experiments, including TROSY and methods that take advantage of favorable relaxation properties. It describes how differential relaxation of spin states can be used to improve resolution and sensitivity. Specific experiments mentioned include TROSY, ZQ-TROSY, CoCaIn for measuring RDCs, and using polychromatic pulses for polarization transfer in large molecules.

1. Spin-state diagramm and I / S correlation spectrum of a spin ½ IS spin system T ransverse R elaxation O ptimized S pectroscop Y ( TROSY )

2. T ransverse R elaxation O ptimized S pectroscop Y ( TROSY ) Prof. K. Pervushin, BioNMR group , LPC, D-CHAB, ETH Zürich

3. An overview

4. Chemical shift correlations in protein backbone spin systems using TROSY: 120 kDa Aldolase

5. Spin-state diagramm and I / S correlation spectrum of a spin ½ IS spin system

7. Evolution of density operator for static molecule

8. Hamiltonian representation as product of spherical harmonics and irreducible tensors of 2 nd rank

10. Evolution of an ensemble of spins

11. Spin relaxation

12. DD/DD interference –reducing relaxation

13. DD/DD interference –enhancing relaxation

14. Chemical Shift Anisotropy (CSA) relaxation H DD (I,S) =  I  S h/r 3 [ 2IzSz – IxSx  IySy ] H CS (I) =  I [  1 IzHz +  2 (IxHx + IyHy)] H CS (I) = 1/3  I [(  1 + 2  2 ) H*I + (  1 -  2 ) ( 2IzHz  IxHx  IyHy )]

15. DD/CSA interference –enhancing relaxation

16. DD/CSA interference –reducing relaxation 

17. CSA/CSA interference –enhancing relaxation

18. CSA/CSA interference –reducing relaxation

19. TROSY effect as function of B 0

21. Spin-state diagramm and I / S correlation spectrum of a spin ½ IS spin system

22. Spin-state diagramm and I / S correlation spectrum of a spin ½ IS spin system

23. Fundamental bounds associated with polarization/coherence transfer imposed by qunatum spin dynamics   C 1. Maximum transfer bound, U 2. Minimal spin-evolution time required for the transfer,  min 3. Suppression of spurious transfers,   Q 4. Combined use of more source operators,   C 5. Complexity of pulse sequence

24. Computer-based design of NMR (near) optimal experiments …  …

25. Computer-based design of NMR (near) optimal experiments

26. Use of MD simulations in the space of pulse sequence variables for constructing of optimal NMR experiments

27. Statistics of computer-based design of Methyl TROSY experiments  = 1/J

28. Complexity versus efficiency of NMR experiments  = 1/J …  … n

29. TROSY (ST2-PT) of Pervushin et al. is theoretically optimal (!!!)  = 1/J (minimal) b/bmax=100% n =2 Source: 1 H+ 15 N No spurious transfers

30. TROSY of Kay et al. is theoretically optimal  = 1/J (minimal) b/bmax=100% n =2 Source: 1 H+ 15 N No spurious transfers

31. ZQ-TROSY of Pervushin et al. is theoretically optimal  = 0.5/J (minimal) b/bmax=100% n =1 Source: 1 H+ 15 N No spurious transfers

32. 1 H- 15 N RDCs measurements with COCAIN TROSY

33. Time-, magnetization source- and transfer efficiency-optimal CoCaIn experiment: theoretically optimal pulse sequence  I z S   1/2 I  ( E + 2 S z ) = 1/2 I    I z S    1/2 I  ( E  2 S z ) = 1/2 I    = 0.5/J (minimal) b/bmax=100% n =1 Source: 1 H+ 15 N No spurious transfers

34. Time-, magnetization source- and transfer efficiency-optimal CoCaIn experiment: Spectra

35. RDCs in methyl groups 13 C + 1 H x -> 1 H    1 H  

36. Construction of optimal NMR experiments

37. Measurements of 1 H- 1 H RDCs in methyl groups

39. The primate erythrocyte/immune complex clearing mechanism

40. Human complement receptor type 1 (CR1)

41. INEPT-based HSQC of 220 kDa CR1/C3b complex  2 ( 1 H) [ppm]  1 ( 15 N) [ppm]

42. Differential driving of the manifolds I  and I  by selective rf-pulse I z = I  z  + I  z -> I  z   I  z = 2 I z S z I  i  = I i  (1/2 E + S z ) I  i  = I i  (1/2 E  S z ) I  z  I  z

43. Excitation profile of polychomatic pulse

44. Polychomatic pulse wave-form and spin trajectory

45. Polarization transfer using polychromatic irradiation  2 ( 1 H) [ppm]  1 ( 15 N) [ppm] CRINEPT POLY-C

46. PC-SPI spectra of free CR1 and CR1/C3b complex

47. CR1/C3b complex CR1 22 kDa CR1/C3b complex 220 kDa