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3D_and_Field_Gradients_10

3D_and_Field_Gradients_10 - Extensions to 3D and Efficiency...

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Extensions to 3D and Efficiency Improvement Using Pulsed Field Gradients BCMB/CHEM 8190
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2D NMR spectra can get very crowded 2D NOESY of Staph- Nuclease 156 AA Solution: go to 3D
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3D experiments used for sequential resonance assignments: Can detect C α in 3D by INEPT transfer from 13 C to 15 N then from 15 N to 1 H. Sequential connections from: HNCA plus HN(CO)CA 1D 2D 3D f3 ( 1 H) f1 ( 13 C) f2 ( 15 N) HN(CO)CA
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Strategy for Constructing 3D Experiments: Combine various 2D Experiments 4D combine more? Cavanagh et al 1996
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3D TOCSY-HSQC Cavanagh et al 1996
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TOCSY-HSQC for 15 N-Labeled Ubiquitin Cavanagh et al 1996
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Limitations for Multidimensional NMR Experiments can be time consuming For 3D we collect t1 and t2 a point at a time – 32 points in t1 and 32 point in t2 require 1024 spectra – with 16 pulse phase cycle requires 9 hrs at 2s rep rate 4D with another 32 point t3 dimension would be 12 days One solution: GFT: Szyperski T, Atreya HS. Mag.
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