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大肠杆菌中产生的心房利钠因子原激素的表达、纯化及体内活性

Expression, purification, and in vivo activity of atrial natriuretic factor prohormone produced in Escherichia coli.

作者信息

Gierse J K, Olins P O, Devine C S, Marlay J D, Obukowicz M G, Mortensen L H, McMahon E G, Blaine E H, Seetharam R

机构信息

Corporate Research and Development, Biological Sciences, Monsanto Company, Chesterfield, Missouri 63198.

出版信息

Arch Biochem Biophys. 1989 Jun;271(2):441-6. doi: 10.1016/0003-9861(89)90294-4.

Abstract

Atrial muscles of the heart are known to produce polypeptide hormones called atrial natriuretic factors (ANF) which have potent diuretic and hypotensive action. These hormones are synthesized as a larger protein precursor called pro atrial natriuretic factor or proANF which contains the biologically active ANF sequences at its C-terminus. Rat proANF (representing amino acids -1 to 128 of the coding sequence) was expressed in a soluble form in Escherichia coli. A simple purification procedure was developed which consists of boiling E. coli cell extracts in 1 M acetic acid and subjecting the supernatant to reversed-phase HPLC. The effect of intravenous administration of the purified recombinant proANF on mean arterial blood pressure was examined. The displacement dose-response curves obtained demonstrated that proANF exhibits similar, albeit less potent, physiological activity than ANF.

摘要

已知心脏的心房肌会产生一种名为心房利钠因子(ANF)的多肽激素,它具有强大的利尿和降压作用。这些激素是以一种名为前心房利钠因子或proANF的更大蛋白质前体形式合成的,该前体在其C末端包含具有生物活性的ANF序列。大鼠proANF(代表编码序列的第 -1 至 128 个氨基酸)在大肠杆菌中以可溶形式表达。开发了一种简单的纯化程序,包括在 1 M 乙酸中煮沸大肠杆菌细胞提取物,并将上清液进行反相高效液相色谱分析。检测了静脉注射纯化的重组proANF对平均动脉血压的影响。所得的置换剂量 - 反应曲线表明,proANF表现出与ANF相似的生理活性,尽管效力较弱。

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