Pre- heteronuclear biomolecular NMR works

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Article [1] presents nearly complete 1H assignment of 129 residue hen egg lysozyme based on Phase-sensitive 2D NOESY, Phase-sensitive COSY and RELAY and Double RELAY spectra culminating a 15 year effort of assigning lysozyme exclusively by 1H NMR. The article presents a detailed illustrated explanation of the sequential assignment process. Spectra were recorded at 35oC. Samples were 7 mM, pH=3.8. Total of four samples were analyzed:

  1. in 90% H2O/10% D2O,
  2. fully D2O exchanged sample repared by heating at 80oC for 10 min
  3. partially D2O exchanged sample prepared by dissolving protein in D2O at 35oC; this sample retained slowly exchanging amide protons
  4. reverse-exchanged sample prepared by dissolving lyophylized fully D2O exchanged sample in 80% H2O/20% D2O. Solid sample was first dissolved in D2O then a proportional amount of H2O was added; this sample was used to detect quickly exchanging amide protons


  1. Redfield, C and Dobson, CM. Sequential proton NMR assignments and secondary structure of hen egg white lysozyme in solution. Biochemistry 27(1):122--136, 1988. BibTeX [red98]
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