LECTURE 5 Spin systems in proteins B ■ H H 1 \ / H-N H c-H I I / H-C-C-C-H II V H 0 = C H /C-H H~N H H H I I H-C-C-H I I 0 = C H H |_| 1 \ / H-N H c-C II// W H-C-C-C C-H I I \ / 0 = C H /C = CX 1 H H H- N i H i H- I c-i I C i 0 = i c 1 i H H- 1 N i H i H- I c-i I C i 0 = i c 1 i H H- 1 N i H i H- I c-i I C i 0 = i c i H H H \ / . C-H / \ H H H H \ / c-c // w C-H ■ H H 1 \ / H-N H c-H I I / H-C-C-C-H / \ H H 1 H A" H H i H H I C i -H i H H H H \ / c- C 1 // w C -c c | \ / H p= c\ H H H- N i H i H- I c-i I C i 0 = i c 1 i H H- 1 N i H i H- I c-i I C i 0 = i c 1 i H H- 1 N i H i H- I c-i I C i 0 = i c i H H H \ / . C-H / \ H H H H \ / c-c // w C-H / \ H H H- N i H i H- I c-i I C i 0 = i c 1 i H H- 1 N i H i H- I c-i I C i 0 = i c 1 i H H- 1 N i H i H- I c-i I C i 0 = i c i H H H \ / . C-H / \ H H H H \ / c-c // w C-H / \ H H Presaturation WATERGATE 3.5e+06 3.0e+06 h 2.5e+06 h 2.0e+06 h 1.5e+06 h 1 .Oe+06 5.0e+05 O.Oe+00 h -5.0e+05 36.9 °C 42.4 °C 47.9 °C 53.4 °C 58.9 °C 64.5 °C 70.0 °C 75.6 °C 81.1 °C 86.7 °C 92.3 °C 97.9 °C 103.5 °C 109.1 °C 114.7 °C 1.2 1.0 0.8 0.6 0.4 0.2 5(H) / ppm 3.5e+06 3.0e+06 h 2.5e+06 h 2.0e+06 h 1.5e+06 h 1.0e+06 h 5.0e+05 h O.Oe+00 h -5.0e+05 36.9 °C 42.4 °C 47.9 °C 53.4 °C 58.9 °C 64.5 °C 70.0 °C 75.6 °C 81.1 °C 86.7 °C 92.3 °C 97.9 °C 103.5 °C 109.1 °C 114.7 °C 1.2 1.0 0.8 0.6 0.4 0.2 5(H) / ppm 1H-15N HSQC S(15N)/ppm J_I_L * I ♦ •« "O "O 3 HNCA, a triple resonance experiment 15N 13q« 13CO J_ J_ y |4Jhn|4JHn| 1 1 — — hi |4JHn|4Jhn I ta 1 1 ± -JL.tij 4Jnc |4JNC2 / III 4JNc 1 1 y 4Jnc I 1 I1 GARP 1 1 |t2| 1 1 1 amino acid_ G A C, E,H,K,M,P,Q,R,V,W D, F,I,L,N,Y,C (S-S) S,T NMR Cf3 chemical shift G no A < 25 ppm O 25 to 36 ppm X 36 to 45 ppm U 55 to 70 ppm PACDEFGHIKLMNQRSTVWY 0A0X0XG0X0X0X00UU00X a = HNCACB b = CBCA(CO)NH abababababababababab Ši í'-'.'-.i.'^áteäí (BS O O C\l o o KT O a O to Y80 S8J V82 T83 Y84 085 G86 F87 N88 T89 F90 Zidek et a I., Biochemistry 1999