High-sensitivity sequence analysis of peptides and proteins by 4-NN-dimethylaminoazobenzene 4'-isothiocyanate.
A manual high-sensitivity sequencing method is described, in which 4-NN-dimethylaminoazobenzene 4'-isothiocyanate is used for the stepwise degradation of amino acid residues from the peptides. The 4-NN-dimethylaminoazobenzene 4'-thiazolinones of amino acids that were released, after conversion into their thiohydantoin derivatives, were identified by t.l.c. on polyamide sheets. This new method is simple and sensitive, and requires only 2-10nmol of peptides or proteins for extended sequence analysis. The method was tested on the sequence analysis of a hexapeptide (Leu-Trp-Met-Arg-Phe-Ala), bradykinin, glucagon and native lysozyme. Results show that the proposed procedure is a sensitive method for the sequence determination of short peptides as well as for the partial sequence determination of intact proteins.
D000595:Amino Acid Sequence; D008722:Methods; D010455:Peptides / Q000032:analysis*; D011506:Proteins / Q000032:analysis*; D013861:Thiocyanates / Q000138:chemical synthesis; D004124:p-Dimethylaminoazobenzene / Q000031:analogs & derivatives* / Q000138:chemical synthesis
D010455:Peptides; D011506:Proteins; D013861:Thiocyanates; D004124:p-Dimethylaminoazobenzene