Abstract
4-N,N-Dimethylaminoazobenzene 4′-isothiocyanate degradations permit sensitive and fast manual sequence analysis, but assignments of some residues are difficult. Hydrophobic residues, especially leucine and isoleucine, are badly resolved on polyamide thin-layer chromatography. Differently colored by-products have been described before for a few labile residues, but it is now shown that most residues can give rise to characteristic by-products. These have different colors and chromatographic properties, depending on the nature of the parent residue. Thus, two to three sets of spots characterize each residue, giving multiple identification with increased reliability. Although variable and dependent on chemicals and conditions, by-products are often prominent after conversion with 50% trifluoroacetic acid, and can be utilized to improve the identifications.
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Abbreviations used are DABITC: 4-N,N-dimethylaminoazobenzene 4′-isothiocyanate; DABTH: dimethylaminoazobenzene thiohydantoin; DABTC: dimethylaminoazobenzene thiocarbamoyl; DABTZ: dimethylaminoazobenzene thiazolinone; dansyl: 5-dimethylaminonaphthalene 1-sulfonyl; HPLC: high-performance liquid chromatography; PTH: phenylthiohydantoin; TFA: trifluoroacetic acid.
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von Bahr-Lindström, H., Hempel, J. & Jörnvall, H. By-products as an aid in residue identification during peptide sequence analysis with dimethylaminoazobenzene isothiocyanate. J Protein Chem 1, 257–262 (1982). https://doi.org/10.1007/BF01025003
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DOI: https://doi.org/10.1007/BF01025003