Boarder M R, Lockfeld A J, Barchas J D
J Neurochem. 1982 Jul;39(1):149-54. doi: 10.1111/j.1471-4159.1982.tb04713.x.
A radioimmunoassay specific for the COOH-terminus of Met-enkephalin [Arg6, Phe7] and a separate assay specific for the COOH-terminus of Met-enkephalin are described. Immunoreactivity by these two assays was compared in bovine caudate and bovine chromaffin granule preparation after Sephadex G75 chromatography in 50% acetic acid. When the assays were applied to the chromatography fractions of the bovine caudate extract, the majority of the immunoreactivity was found in the fractions corresponding to the heptapeptide and the pentapeptide respectively. When the chromaffin granule chromatography fractions were assayed, both of the radioimmunoassays showed that most reactivity was in several peaks in the larger molecular weight fractions. The major peak for the Met-enkephalin [Arg6, Phe7] assay had an apparent molecular weight of 2800, while with the Met-enkephalin assay the dominant peak of immunoreactivity had an apparent molecular weight of 10,000. The presence of authentic Met-enkephalin [Arg6, Phe7] in both caudate and chromaffin granule extracts was confirmed by reverse-phase chromatography of the previously sized fractions. It appears then that the processing of precursors of opioid peptides is directed, in the caudate, to the synthesis and storage of the enkephalins and of Met-enkephalin [Arg6, Phe7]; in the adrenal medulla the major products of precursor processing are a variety of polypeptides of larger sizes.
本文描述了一种针对甲硫氨酸脑啡肽[精氨酸6,苯丙氨酸7]羧基末端的放射免疫分析方法以及另一种针对甲硫氨酸脑啡肽羧基末端的独立分析方法。在50%乙酸中经葡聚糖凝胶G75层析后,对牛尾状核提取物和牛嗜铬颗粒制剂进行了这两种分析方法的免疫反应性比较。当将这些分析方法应用于牛尾状核提取物的层析级分时,大部分免疫反应性分别在对应于七肽和五肽的级分中被发现。当对嗜铬颗粒层析级分进行分析时,两种放射免疫分析方法均显示大部分反应性存在于较大分子量级分的几个峰中。甲硫氨酸脑啡肽[精氨酸6,苯丙氨酸7]分析方法的主要峰的表观分子量为2800,而甲硫氨酸脑啡肽分析方法的主要免疫反应性峰的表观分子量为10000。通过对预先分级的级分进行反相层析,证实了尾状核提取物和嗜铬颗粒提取物中均存在真实的甲硫氨酸脑啡肽[精氨酸6,苯丙氨酸7]。由此看来,在尾状核中,阿片肽前体的加工过程是朝着脑啡肽和甲硫氨酸脑啡肽[精氨酸6,苯丙氨酸7]的合成与储存进行的;而在肾上腺髓质中,前体加工的主要产物是多种较大尺寸的多肽。