Steele P A, Turner C A, Murphy R
Department of Physiology, School of Medicine, Flinders University, Bedford Park, Australia.
Neuropeptides. 1989 Apr;13(3):207-13. doi: 10.1016/0143-4179(89)90094-2.
Guinea-pig ileum was dissected and the mucosa, submucosa and external musculature extracted with aqueous acetic acid for measurement of four prodynorphin-derived peptides, namely dynorphin A 1-8, dynorphin A 1-17, dynorphin B, and alpha-neoendorphin. The peptide-like immunoreactive material extracted from the external musculature was characterized by multi-dimensional chromatographic analysis and compared to synthetic porcine standards. The chromatographic methods utilized were: reversed-phase high performance liquid chromatography (RP-HPLC), using two different eluants; cation exchange high performance liquid chromatography (CE-HPLC) and gel filtration chromatography. The dynorphin A 1-8-like immunoreactive material was homogeneous and coeluted with the standard in all chromatographic modes. The dynorphin A 1-17-like and dynorphin B-like immunoreactive material was heterogeneous but showed a peak that coeluted with synthetic standard in all chromatographic modes. The alpha-neoendorphin-like immunoreactive material also appeared to be heterogeneous with the major component on CE-HPLC coeluting with the synthetic peptide standard while the major component on RP-HPLC eluted differently. It was concluded that the guinea-pig ileum contains immunoreactivity for peptides derived from all coding regions of the prodynorphin gene and that these peptides may be present in multiple immunoreactive forms.
解剖豚鼠回肠，用醋酸水溶液提取黏膜、黏膜下层和外部肌肉组织，以测定四种前强啡肽衍生肽，即强啡肽A 1-8、强啡肽A 1-17、强啡肽B和α-新内啡肽。从外部肌肉组织中提取的肽样免疫反应物质通过多维色谱分析进行表征，并与合成猪标准品进行比较。所采用的色谱方法包括：反相高效液相色谱（RP-HPLC），使用两种不同的洗脱液；阳离子交换高效液相色谱（CE-HPLC）和凝胶过滤色谱。强啡肽A 1-8样免疫反应物质是均一的，并且在所有色谱模式下均与标准品共洗脱。强啡肽A 1-17样和强啡肽B样免疫反应物质是异质的，但在所有色谱模式下均显示出与合成标准品共洗脱的峰。α-新内啡肽样免疫反应物质似乎也是异质的，CE-HPLC上的主要成分与合成肽标准品共洗脱，而RP-HPLC上的主要成分洗脱方式不同。得出的结论是，豚鼠回肠含有对源自前强啡肽基因所有编码区的肽的免疫反应性，并且这些肽可能以多种免疫反应形式存在。
