Amino acid sequence determination by g.l.c.-mass spectrometry of permethylated peptides. Optimization of the formation of chemical derivatives at the 2-10 nmol level

Authors:
  • KRK Rose
    K Rose
  • MSM G Simona
    M G Simona
  • ROR E Offord
    R E Offord
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Abstract
A new technique is described that permits the permethylation of acylated peptides at the 2-10 nmol level. The presence of up to 400 micrograms of sodium dodecyl sulphate per sample does not affect the reaction yields. The technique, which is a miniaturization of the widely used methyl iodide/dimethylsulphinyl carbanion procedure, employs a layer of hexane to exclude moisture and oxygen from the reaction mixture. Analysis of the peptide derivatives by combined g.l.c.-mass spectrometry permits amino acid sequence information to be obtained. In addition to studies of digests of a model substrate (glucagon), the new permethylation technique has been used to identify a peptide of interest from a digest of a cytochrome and to define the N-termini of two proteins at the 5 nmol level.

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