Protein/Protein and Protein/Ligandinteractions in large and dynamicallydisordered systems studied by NMR insolution

1. NMR structure/dynamics of apo- and holo-forms of heme chaperone ccmE

2. Protein/Protein and Protein/Ligand interactions in large and dynamically disordered systems studied by NMR in solution Prof. K. Pervushin, BioNMR group , LPC, D-CHAB, ETH Zürich

4. Chemical shift correlations in protein backbone spin systems using TROSY

5. Remodelling of outer membrane protein A A NMR conformer of the N-terminal domain of OmpA A BBP NMR structure exterior outer membrane periplasmic space EF-hand loop III Tb 3+ Thr Ser Asp Lys Asp Gly Asn Gly Tyr Ile Ser Ala Ala Glu Ala Ser

6. NMR structure/dynamics of apo- and holo-forms of heme chaperone ccmE

7. Role of flexible C-terminal 15 amino acids of 44 kDa BsCM in catalysis Endo -oxabicyclic transition state analog, TSA Putative transition state

8. Refolding of HTH DBP protein in 6 M Urea by Hofmeister reagents HTH in 6 M Urea unfolded HTH in 6 M Urea, 1.7 M NaCl native structure refolding HTH in 6 M Urea unfolded HTH in 6 M Urea, 0.5 M NaTFA distorted structure, molten globule refolding

9. Engineered monomeric chorismate mutase lacking a preorganized structure 1

11. The primate erythrocyte/immune complex clearing mechanism

12. Human complement receptor type 1 (CR1)

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

14. Fundamental bounds associated with polarization/coherence transfer imposed by quantum 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

15. Definition of the optimization problem   C U H =  J  I z S z +  x (t) I x +  y (t) I y

16. Definition of the optimization problem for isolated 2 spin ½ system S I DD( IS ), CSA( S ) and CSA( I ) interactions max (Khaneja et al, PNAS, 2003, 100, 13162)

17. 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

18. Excitation profile of polychomatic pulse

19. Polychomatic pulse wave-form and spin trajectory

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

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

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

24. 54 kDa „moonlight“ chaperone with PPIase activity 65 Å Substrate

25. 54 kDa „moonlight“ chaperone with PPIase activity

26. 15 N relaxation measurements of free FkpA at 600 MHz

27. 15 N relaxation measurements with FkpA at 600 MHz

28. 1 H- 15 N RDCs measurements in the presence of Pf1 phages

29. Histogramm of RDCs values in two media C 12 E 5 / hexanol/H 2 O L  n -Alkyl-poly(ethylene glycol)/ n -alkyl alcohol and glucopone/ n -hexanol mixtures Phages Pf1

30. RDCs values in Pf1 medium

31. RDCs values in Pf1 medium

32. A schematic model of intramolecular dynamics in FkpA

33. Chemical shift changes by complex formation with (1) reduced and carboxymethylated bovine  -lactalbumin, (2) RNAse AS

34. Chemical shifts mapping

35. Equilibrium binding of FkpA to substrates: (1) reduced and carboxymethylated bovine  -lactalbumin, (2) RNAse AS K d = 540  m

36. Protein Quality Control in the ER

37. Substrates recognized by GT RNase B RNase BS RNase B S protein alkylated RNase B - + - GT: RNase BS” S peptide 15-mer scrambled RNase B small glyco- peptides + - - -

39. RNase A 15 N- 1 H HSQC RNase A: complete assignment is available

41. Chemical Shift Difference between S protein and RNase A

42. Fast Amide Proton Exchange

43. 15 N-Relaxation measurements

44. R ex by cross-correlated relaxation 0 20 40 60 80 100 120 20 30 40 50 60 70 80 90 100 110 120 Residue Number R 2 , s -1 R 2 R 2 - R ex fr. CCR

45. Concentration Scan 1.06 mM

46. Concentration Scan 0.2 mM

47. Concentration Scan 0.08 mM

48. Ratio between peak volumes corresponding to oligomerization states of RNAse AS R = V oligomeric /V monomeric RNAse AS [mM]

49. Lys 60 Delution Chaperone

50. Gln 65 Delution Chaperone

51. Leu 91 Delution Chaperone

52. Lys 95 Delution Chaperone

53. Conformational dynamics in S Protein S Protein N S Protein U k u k f [S Protein] n >30ms ~80 Hz   k c

54. 15 N relaxation measurements of FkpA/S-protein complex at 600 MHz

55. 15 N relaxation measurements of free FkpA at 600 MHz

56. A „mother‘a arms“ model of chaperone activity of FkpA

57. Thanx a lot! Alexander Eletski Prof. Donald Hilvert Beat Vögeli Prof. Linda Thöny-Meier Dr. Osvaldo Moreira Prof. Andreas Plückthun Kaifeng Hu Dr. Helena Kovac (Bruker AG) Alexander Kienhoffer Dr. Maria Johansson Simon Alioth Katherina Vamvaca Krystina Bromek Dr. Donghan Lee SNF and ETH for financial support Prof. Paul Barlow Prof. Ari Helenius Dr. Christiana Ritter