Sensitization of Edman amino acid derivatives using the fluorescent reagent, 4-aminofluorescein.
The ability to analyze amino acid derivatives at the femtomole level is one of the most interesting challenges in the field of protein microsequencing. 2-Anilino-5-thiazolinone amino acids, obtained by Edman degradation, were quantitatively derivatized with fluorescent primary amines. The most fluorescent reagent tested was 4-aminofluorescein. The amino acid derivatives sensitized with this reagent were separated using reversed-phase high-performance liquid chromatography and identified at the 100 attomole level. Incorporation of this method into the operation of a conventional automated sequencer is also described.
Tsugita
A
A
Research Institute for Biosciences, Science University of Tokyo, Chiba.
Kamo
M
M
Jone
C S
CS
Shikama
N
N
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis*; D000814:Aniline Compounds / Q000032:analysis; D055598:Chemical Phenomena; D002621:Chemistry; D002851:Chromatography, High Pressure Liquid; D005452:Fluoresceins; D005453:Fluorescence; D006863:Hydrogen-Ion Concentration; D008722:Methods; D013844:Thiazoles / Q000032:analysis; D013859:Thiocarbamates / Q000032:analysis
D000596:Amino Acids; D000814:Aniline Compounds; D005452:Fluoresceins; D013844:Thiazoles; D013859:Thiocarbamates; C061079:4-aminofluorescein