RDCs_10 - Residual Dipolar Couplings BCMB/CHEM 8190 Recent...

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Residual Dipolar Couplings BCMB/CHEM 8190
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Recent Reviews • Prestegard, A-Hashimi & Tolman, Quart. Reviews Biophys. 33 , 371-424 (2000). • Bax, Kontaxis & Tjandra, Methods in Enzymology , 339 , 127-174 (2001) • Tolman, Curr. Opin. Struct. Biol . 11 , 532-539 (2001) • Prestegard, Bougault & Kishore, Chemical Reviews , 104 , 3519-3540 (2004) • Lipsitz & Tjandra, Ann. Rev. Biophys. Biomol. Struct ., 33 , 387-413 (2004) • Fushman et al., Prog. NMR Spect . 44 , 189-214 (2004)
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The Dipolar Interaction Between Two Spins 15 N 1 H B 0 r HZ NZ 2 3 I I 2 1 θ 3cos r C = D Brackets denote averaging – goes to zero without partial orientation
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Inducing Order Using Liquid Crystalline Media B 0
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Measurement of Dipolar Couplings – Coupled HSQC Isotropic Aligned J J + D
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Polyacrylamide Gels another alignment medium Yizhou Liu, J. Prestegard J. Biomol NMR , submitted
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31 2 3 2 cos θ φφ ρ δ = ⋅⋅⋅ ⋅⋅ × ij kl k l Order Matrix Analysis x y φ z ρ z θ ρ x z
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Finding a Principal Order Frame S xx S xy .. S yx S yy .. .. .. .. =A S x’x’ S y’y’ S z’z’ A -1
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Strategy for Protein Fold Determination • Express 15 N labeled protein • Identify secondary elements •Assign backbone resonances •Orient protein in LC medium •Collect residual dipolar data • Orient individual elements • Assemble protein fold
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Dipolar Interaction Vectors In an Idealized α -Helix 15 N Labeling Only
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Data Used in ACP Fold Determination A. Dipolar couplings in α -helices Couplings (Hz) Helix 1 Helix 2 Helix 3 Amide Couplings I3 1.4 L37 -2.6 Q66 8.2 (N i -H N i ) E4 0.4 D38 1.6 A67 7.7 E5 3.4
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RDCs_10 - Residual Dipolar Couplings BCMB/CHEM 8190 Recent...

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