Lecture 19 - X RAY NMR

Distance between bonded h atoms three chemical bonds

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Fingerprint of a.a. – Distance between BONDED H atoms ( three chemical bonds, I.e. within the same a.a.) • NOESY (NOE, Nuclear Overhausser Effect) – Distance between H atoms close together in space (
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Interpretation Sequence Specific Assignment or Sequential assignment 1. COSY – Cross peak unique for each amino acid, “fingerprint” Which a.a. in sequence? 2. NOESY a. interactions in space b. interactions of residues that are sequentially adjacent (i and i+1) 3. Amino Acid Sequence 4. Distance Constraints: for H atoms in i to H atoms in j { Cross-peaks
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Interpretation Sequence Specific Assignment or Sequential assignment 1. COSY – Cross peak unique for each amino acid, “fingerprint” Which a.a. in sequence? 2. NOESY a. interactions in space b. interactions of residues that are sequentially adjacent (i and i+1) 3. Amino Acid Sequence 4. Distance Constraints: for H atoms in i to H atoms in j { Cross-peaks
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Interpretation cont… 5. Structure Refinements - computer modeling - No unique structure but different structures that are compatible with data (ambiguity)
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By Combination of x-ray and NMR • Structure Determination • Complementary • In general agreement (except minor discrepancies especially loops) •Bo th requ
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Determining Protein Structure • X-Ray Diffraction – Interactions of x-rays with electrons in molecules in a crystal • NMR- Nuclear Magnetic Resonance – Interactions of magnetic field (external) with the intrinsic magnetic properties of atomic nuclei which possess a spin angular momentum COMPLEMENTARY
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Distance between BONDED H atoms three chemical bonds Ie...

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