牛心房利钠因子的纯化与序列测定

Purification and sequence determination of bovine atrial natriuretic factor.

作者信息

Ong H, McNicoll N, Lazure C, Seidah N, Chrétien M, Cantin M, De Léan A

出版信息

Life Sci. 1986 Apr 7;38(14):1309-15. doi: 10.1016/0024-3205(86)90425-x.

Abstract

We report the purification and the sequence determination of bovine atrial natriuretic factor (ANF) in acid extracts of bovine atrial appendages. The monitoring of the activity along the purification steps was performed with a radio-receptor assay using bovine adrenal cortex membranes sites and 125I ANF. Bovine ANF was separated by carboxymethyl agarose gel chromatography from catecholamines and major protein contaminants. It behaved as a 3 K dalton peptide on Sephadex G-50. The active fractions were then subjected to high performance liquid chromatography (HPLC) using a sulfopropyl cation exchange column. Subsequent purification steps by reverse phase mode on Ultrapore RPSC, Vydac TP 218 and uBondapak using acetonitrile gradients led to the obtention of a pure fraction which amino acid sequence was identical to that for human ANF. This confirms the high degree of homology of ANF structure among mammalian species and advocates the use of the radio-receptor assay based on bovine adrenal receptor for measuring human ANF.

摘要

我们报道了从牛心耳酸提取物中纯化和测定牛心房利钠因子（ANF）的过程。在纯化步骤中，使用牛肾上腺皮质膜位点和125I标记的ANF通过放射受体测定法监测活性。牛ANF通过羧甲基琼脂糖凝胶色谱法与儿茶酚胺和主要蛋白质污染物分离。在葡聚糖G - 50上它表现为一种3千道尔顿的肽。然后使用磺丙基阳离子交换柱对活性级分进行高效液相色谱（HPLC）。随后在Ultrapore RPSC、Vydac TP 218和uBondapak上使用乙腈梯度通过反相模式进行进一步纯化步骤，得到了一个纯级分，其氨基酸序列与人ANF的序列相同。这证实了哺乳动物物种间ANF结构的高度同源性，并提倡使用基于牛肾上腺受体的放射受体测定法来测量人ANF。