Proenkephalin A and proopiomelanocortin peptides in human cerebrospinal fluid.
Precursors to beta-endorphin (BE) and methionine enkephalin (ME), and proteolytic enzymes that cleave those BE and ME precursors to BE and ME, were determined in several milliliters of human cerebrospinal fluid. Endogenous peptides were purified by reversed-phase high-performance liquid chromatography (HPLC), and were detected with radioreceptor assay (RRA), radioimmunoassay, and mass spectrometry (MS). Total opioid receptor activity measurements and the profile of HPLC-receptor activity of human CSF samples were both used to monitor neuropeptide metabolism. MS data linked the molecular ion of ME to a unique fragment ion. A later-eluting fraction (84 min) in a 90-min HPLC gradient appeared in all HPLC-RRA profiles, contained opioid receptor activity that displaced [3H]etorphine, and the quantitative and qualitative patterns of opioid receptor activity in those profiles both changed within the few minutes that elapsed between acquiring the first and second cerebrospinal fluid samples. That 84-min fraction contained precursors to opioid peptides and was fractionated further with a more shallow 120-min HPLC gradient into three sections that displayed delta-opioid receptor-preferring activity, using [3H]ME as ligand. These three sections were hydrolyzed separately with human cerebrospinal fluid as the source for endogenous neuropeptides to yield products that correlated to immunoreactive BE in section I and immunoreactive ME in section III.
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Charles B. Stout Neuroscience Mass Spectrometry Laboratory, University of Tennessee, Memphis 38163.
Wood
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Desiderio
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D002851:Chromatography, High Pressure Liquid; D004744:Enkephalin, Methionine / Q000134:cerebrospinal fluid; D004745:Enkephalins / Q000134:cerebrospinal fluid*; D006801:Humans; D006868:Hydrolysis; D013058:Mass Spectrometry; D011333:Pro-Opiomelanocortin / Q000134:cerebrospinal fluid*; D011498:Protein Precursors / Q000134:cerebrospinal fluid*; D011863:Radioimmunoassay; D011869:Radioligand Assay; D011957:Receptors, Opioid / Q000032:analysis / Q000187:drug effects; D001615:beta-Endorphin / Q000134:cerebrospinal fluid
D004745:Enkephalins; D011498:Protein Precursors; D011957:Receptors, Opioid; C029992:proenkephalin; D004744:Enkephalin, Methionine; D001615:beta-Endorphin; D011333:Pro-Opiomelanocortin