Characterization of opioid peptides in the rat stomach.
A combination of several chromatographic and assay systems was used to characterize the opioid peptides in rat stomach extracts. Partial purification of opioid material in acetic acid extracts of the corpus plus antrum regions of the rat stomach was carried out by gel filtration chromatography on Sephadex G-50, followed by adsorption onto Amberlite XAD-2 resin. A single peak in opioid activity was determined by both radioreceptor assay (RRA) and bioassay. By high performance liquid chromatography, this peak was resolved into five distinct components, characterized by RRA and (or) radioimmunoassay, with retention times corresponding to methionine enkephalin (met-enk), leucine enkephalin, met-enk-arg6-gly7-leu8, met-enk-arg6-phe7, and dynorphin 1-13. Closer examination of the dynorphin component revealed the presence of dynorphins 1-17, 1-13, and 1-8. Trypsin digestion of the partially purified (Sephadex G-50 and Amberlite XAD-2 chromatographed) extract resulted in an overall increase in opioid activity, suggesting the presence of larger, possibly precursor forms.
D000818:Animals; D002845:Chromatography; D004399:Dynorphins / Q000032:analysis; D004723:Endorphins / Q000032:analysis* / Q000378:metabolism; D004743:Enkephalin, Leucine / Q000032:analysis; D004744:Enkephalin, Methionine / Q000031:analogs & derivatives / Q000032:analysis; D008297:Male; D011863:Radioimmunoassay; D011869:Radioligand Assay; D051381:Rats; D011919:Rats, Inbred Strains; D013270:Stomach / Q000032:analysis*; D014357:Trypsin
D004723:Endorphins; D004744:Enkephalin, Methionine; D004743:Enkephalin, Leucine; C027511:enkephalin-Met, Arg(6)-Phe(7)-; D004399:Dynorphins; C033453:enkephalin-Met, Arg(6)-Gly(7)-Leu(8)-; D014357:Trypsin