Opioid and tachykinin peptides, and their precursors and precursor-processing enzymes, in human cerebrospinal fluid.
Opioid and tachykinin neuropeptides, which were derived from two biological sources (intact, and released from their corresponding precursors by the action of human cerebrospinal fluid (CSF) neuropeptidases), were characterized in human CSF by using a combination of post-high-performance liquid chromatographic (HPLC) detection techniques. Peptides were separated using gradient and isocratic reversed-phase HPLC. Radioimmunoassay measured immunoreactivity corresponding to several different individual neuropeptides including methionine enkephalin, leucine enkephalin, substance P and beta-endorphin. Commercial enzymes (trypsin, carboxypeptidase B) were used to release methionine- and leucine-enkephalin from precursors. Human CSF also served as a source of endogenous neuropeptidases. Mass spectrometry produced fragment ions that corroborated the amino acid sequence of methionine enkephalin and of substance P derived from both sources (intact, from precursors). These results demonstrated the presence of endogenous intact neuropeptides, several different neuropeptide-containing precursors and appropriate precursor-processing enzymes in human CSF for precursors of methionine enkephalin, leucine enkephalin, beta-endorphin1-31 and substance P.
Liu
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Charles B. Stout Neuroscience Mass Spectrometry Laboratory, University of Tennessee, Memphis 38163.
Dass
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Wood
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Desiderio
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D000595:Amino Acid Sequence; D002623:Chemistry Techniques, Analytical; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004723:Endorphins / Q000134:cerebrospinal fluid*; D004792:Enzyme Precursors / Q000134:cerebrospinal fluid*; D006801:Humans; D013058:Mass Spectrometry; D011863:Radioimmunoassay; D015320:Tachykinins / Q000134:cerebrospinal fluid*
D004723:Endorphins; D004792:Enzyme Precursors; D015320:Tachykinins