A GENERAL PROCEDURE FOR ANALYSIS OF PROENKEPHALIN-B DERIVED OPIOID-PEPTIDES
A GENERAL PROCEDURE FOR ANALYSIS OF PROENKEPHALIN-B DERIVED OPIOID-PEPTIDES
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DOI：
10.1016/0167-0115(85)90032-1
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发表时间：
1985-01-01
影响因子：
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通讯作者：
TERENIUS, L
中科院分区：
文献类型：
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作者：
CHRISTENSSONNYLANDER, I;NYBERG, F;TERENIUS, L
本研究采用胰酶消化和(Leu)脑啡肽-Arg6放射免疫分析法检测阿片肽前激素原脑啡肽B脑啡肽序列的所有可能产物的存在。采用凝胶过滤的方法对不同的前驱体产物按相对分子质量进行分离。分别用高灵敏的强啡肽A和强啡肽B放射免疫分析法监测放散曲线。免疫活性强啡肽A出现3个峰，分子量分别约为1000、2000和5000。强啡肽B部分出现在其他峰(1500、5000和10000道尔顿)。通过测定胰酶消化后凝胶过滤所有组分的脑啡肽-Arg-6免疫活性(Leu)脑啡肽-Arg-6的图谱显示出比强啡肽A和强啡肽B更复杂的图谱，主要峰与强啡肽A和强啡肽B重合，但高相对分子质量区(MW>；5000)也产生高水平的免疫反应(Leu)脑啡肽-Arg-6。
Tryptic digestion followed by radioimmunoassay for (Leu)enkephalin-Arg6 was used in this study as a general method to detect the presence of all possible products containing the enkephalin sequence from the opioid peptide prohormone, proenkephalin B. Tissue extracts of human hypothalamus and pituitary were examined. Gel filtration was used to separate the different precursor products according to MW. The elution profile was also monitored with highly sensitive radioimmunoassays for dynorphin A and dynorphin B, respectively. Immunoreactive dynorphin A appeared in 3 peaks with the approximate MW 1000, 2000 and 5000. Immunoreactive dynorphin B partly occurred in other peaks (1500, 5000 and 10,000 daltons). Profiles obtained by measuring immunoreactive (Leu)enkephalin-Arg-6 in all fractions from gel filtration after trypsin digestion showed a more complex pattern compared to the profiles of immunoreactive dynorphin A and dynorphin B. The major peaks coincided with dynorphin A and dynorphin B but high levels of immunoreactive (Leu)enkephalin-Arg6 were also generated from higher MW regions (MW > 5000).