A new sensitive Edman-type reagent: 4-(N-1-dimethylaminonaphthalene-5-sulfonylamino)phenyl isothiocyanate. Its synthesis and application for micro-sequencing of polypeptides.
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.
Jin
S W
SW
Chen
G X
GX
Palacz
Z
Z
Wittmann-Liebold
B
B
D000595:Amino Acid Sequence; D003619:Dansyl Compounds / Q000138:chemical synthesis*; D007202:Indicators and Reagents / Q000138:chemical synthesis*; D017879:Isothiocyanates; D013058:Mass Spectrometry; D013055:Spectrophotometry, Infrared; D013861:Thiocyanates / Q000138:chemical synthesis*
D003619:Dansyl Compounds; D007202:Indicators and Reagents; D017879:Isothiocyanates; D013861:Thiocyanates; C048325:4-(N-1-dimethylaminonaphthalene-5-sulfonylamino)phenyl isothiocyanate