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Assignment Strategies

Assignment Strategies. O-C-O. O- C -O. H- C -H. O -H. H-C-H. O -H. H- C -H. H- C -H. H-C-H. H-C-H. -N - C a -CO-N - C a -CO-. -N-C a -C O -N-C a -C O -. H. H. H. H. H. H. H. H. Homonuclear Two steps needed Scalar coupling within each amino acid

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Assignment Strategies

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  1. Assignment Strategies O-C-O O-C-O H-C-H O-H H-C-H O-H H-C-H H-C-H H-C-H H-C-H -N-Ca-CO-N-Ca-CO- -N-Ca-CO-N-Ca-CO- H H H H H H H H • Homonuclear • Two steps needed • Scalar coupling within each amino acid • Dipolar coupling between amino acids • Heteronuclear • Build up one atom at a time • Scalar coupling within and between each amino acid; some experiments provide intra- and inter-residue peaks

  2. Pairs of ExperimentsDistinguishes Intra-residue from Inter-residue H H H H H-C-H H-C-H H-C-H H-C-H -N-Ca-CO-N-Ca-CO- -N-Ca-CO-N-Ca-CO- H H H H H H H H • HNCA • H(t1), N(t2), CA(t3) • 2 peaks- Intra + Inter • Mixing from NCa occurs over 1 bond to same Ca and over 2 bonds to adjacent Ca • HN(CO)CA • H(t1), N(t2), CA(t3) • 1 peak- Inter only • Mixing from NCa occurs ONLY over 2 bonds to adjacent Ca

  3. Pairs of ExperimentsDistinguishes Intra-residue from Inter-residue H H H H H-C-H H-C-H H-C-H H-C-H -N-Ca-CO-N-Ca-CO- -N-Ca-CO-N-Ca-CO- H H H H H H H H • HNCACB • H(t1), N(t2), CA and CB(t3) • 4 peaks- intra + inter • Mixing from NCa occurs over 1 bond to same Ca and over 2 bonds to adjacent Ca • HN(CO)CACB • H(t1), N(t2), CA and CB(t3) • 2 peaks- inter only • Mixing from NCa occurs ONLY over 2 bonds to adjacent Ca

  4. Equivalent ExperimentsCan be run in either direction H H H H H-C-H H-C-H H-C-H H-C-H -N-Ca-CO-N-Ca-CO- -N-Ca-CO-N-Ca-CO- H H H H H H H H • HNCACB • H(t1), N(t2), CA and CB(t3) • Mixing from NCa occurs over 1 bond to same Ca and over 2 bonds to adjacent Ca • CBCANH • CB and CA(t1), N(t2), H(t3) • Mixing from NCa occurs over 1 bond to same Ca and over 2 bonds to adjacent Ca

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