针对合成肽的抗血清可识别含高分子量脑啡肽的蛋白质。

Antisera to synthetic peptide recognize high molecular weight enkephalin-containing proteins.

作者信息

Christie D L, Birch N P, Aitken J F, Harding D R, Hancock W S

出版信息

Biochem Biophys Res Commun. 1984 Apr 30;120(2):650-6. doi: 10.1016/0006-291x(84)91305-6.

PMID:6203524

Abstract

Antisera to a synthetic peptide corresponding to the 95-117 sequence of proenkephalin were used to develop a sensitive radioimmunoassay. Gel-filtration of acid extracts of bovine adrenal medulla and purified chromaffin granules revealed that the antisera recognized high molecular weight material (Mr approximately 5,000-30,000). The material in peak I ( Mr 20 ,000-30,000) and peak II (Mr 10,000-20,000) was further purified by immunoaffinity chromatography. Sequential digestion of each of these fractions with trypsin and carboxypeptidase B generated immunoreactive Met-enkephalin. This study demonstrates that antisera against a synthetic peptide cross-react with high molecular weight enkephalin-containing precursors, validating the use of these antisera in studies of enkephalin biosynthesis.

摘要

使用针对脑啡肽原95 - 117序列合成肽的抗血清开发了一种灵敏的放射免疫测定法。对牛肾上腺髓质酸提取物和纯化的嗜铬颗粒进行凝胶过滤显示，抗血清识别高分子量物质（Mr约为5,000 - 30,000）。通过免疫亲和色谱进一步纯化了峰I（Mr 20,000 - 30,000）和峰II（Mr 10,000 - 20,000）中的物质。用胰蛋白酶和羧肽酶B依次消化这些级分中的每一个，产生了具有免疫活性的甲硫氨酸脑啡肽。本研究表明，针对合成肽的抗血清与高分子量含脑啡肽前体发生交叉反应，证实了这些抗血清在脑啡肽生物合成研究中的应用价值。