High-performance liquid chromatography and diode-array detection for the identification of peptides containing aromatic amino acids in studies of endorphin-degrading activity in human cerebrospinal fluid.
Diode-array UV detection has been adapted for analysis of opioid peptides and their metabolic fragments differing in aromatic amino acid content. In combination with high-performance liquid chromatography, the technique allowed a direct and rapid discrimination between peptides containing phenylalanine, tryptophan and tyrosine, or a combination of these residues. Enkephalin fragments with either tyrosine or phenylalanine, or both, were identified after digestion of the pentapeptide with proteolytic activity recovered from human cerebrospinal fluid. The N-terminal tyrosine-containing fragment of dynorphin A was identified after hydrolysis of the peptide by a cerebrospinal fluid endopeptidase. The study was extended to the analysis of some non-opioid peptides. The Tyr1 analogue of delta-sleep-inducing peptide was easily distinguished from the authentic compound with a tryptophan at the N-terminus. Results indicated that the technique was useful for discriminating between dipeptides differing in aromatic residues that were unresolved by high-performance liquid chromatography.
Nyberg
F
F
Pernow
C
C
Moberg
U
U
Eriksson
R B
RB
D000596:Amino Acids / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D004399:Dynorphins / Q000031:analogs & derivatives / Q000134:cerebrospinal fluid; D010450:Endopeptidases / Q000032:analysis; D004723:Endorphins / Q000134:cerebrospinal fluid*; D004743:Enkephalin, Leucine / Q000134:cerebrospinal fluid; D006801:Humans; D015260:Neprilysin; D010446:Peptide Fragments / Q000134:cerebrospinal fluid; D010455:Peptides / Q000302:isolation & purification*; D013056:Spectrophotometry, Ultraviolet
D000596:Amino Acids; D004723:Endorphins; D010446:Peptide Fragments; D010455:Peptides; D004743:Enkephalin, Leucine; D004399:Dynorphins; C039666:dynorphin amide (1-10); D010450:Endopeptidases; D015260:Neprilysin