High-performance liquid chromatographic analysis of synthetic peptides using derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate.
A series of synthetic prothrombin leader sequences (ANKGFLEEX), each having a different C-terminal amino acid, was derivatized with the novel fluorescent-derivatizing agent 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate and analyzed by HPLC. Essentially quantitative yields of derivatized peptides could be routinely obtained with a fast, simple derivatizing procedure. The response was linear over a concentration range of 0.176 to 88.0 nM. Peptides labeled with this highly sensitive derivatizing agent exhibited a mass detection limit in the femtomole range and a concentration detection limit in the picomolar range. Amino acid content of hydrolyzed peptides was carried out using the same reagent in order to evaluate the amino acid composition and to detect any deletion sequences.
De Antonis
K M
KM
University of Rhode Island, Kingston 02881.
Brown
P R
PR
Cohen
S A
SA
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000634:Aminoquinolines; D002219:Carbamates; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000032:analysis* / Q000737:chemistry; D021382:Protein Sorting Signals / Q000032:analysis / Q000737:chemistry; D011516:Prothrombin / Q000737:chemistry
C078178:6-aminoquinolyl-N-hydroxysuccinimidyl carbamate; D000596:Amino Acids; D000634:Aminoquinolines; D002219:Carbamates; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D009842:Oligopeptides; D021382:Protein Sorting Signals; D011516:Prothrombin