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A new sensitive Edman‐type reagent: 4‐( N ‐1‐dimethylaminonaphthalene‐5‐sulfonylamino)phenyl isothiocyanate
Author(s) -
Jin Shan-Wei,
Chen Gui-Xiang,
Palacz Zbigniew,
Wittmann-Liebold Brigitte
Publication year - 1986
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(86)81202-9
Subject(s) - reagent , edman degradation , chemistry , phenyl isothiocyanate , isothiocyanate , chromatography , organic chemistry , peptide sequence , biochemistry , gene
A new fluorescent PITC homologue Edman‐type reagent, 4‐( N ‐1‐dimethylaminonaphthalene‐5‐sulfonylamino) phenyl isothiocyanate (DNSAPITC), was synthesized following a simple three‐step synthetic route. The reagent was crystallized and characterized by thin‐layer chromatography, IR and electron impact mass spectrometry. Reference DNSAPTH‐amino acid derivatives were prepared and a two‐dimensional chromatography system on micro‐polyamide sheets was developed for separating this mixture. On these sheets the sensitivity was 1–5 pmol, by exposure at 366 nm. Model peptides and proteins were subjected to Edman degradations with this new reagent. A similar coupling efficiency and repetitive degradation yield to those of PITC were found with this reagent. The advantages and limitations of this reagent for sensitive microsequencing are discussed.