分离出约32 kDa的牛卵泡液抑制素。

Isolation of bovine follicular fluid inhibin of about 32 kDa.

作者信息

Fukuda M, Miyamoto K, Hasegawa Y, Nomura M, Igarashi M, Kangawa K, Matsuo H

出版信息

Mol Cell Endocrinol. 1986 Jan;44(1):55-60. doi: 10.1016/0303-7207(86)90105-x.

Abstract

Inhibin of about 32 kDa from bovine follicular fluid (bFF) was purified by using chromatographies operated under protein-dissociating conditions, which we have established for our previous purification of porcine follicular fluid (pFF) inhibin. On a gel filtration, bFF inhibin activity was eluted at 3 distinct regions corresponding to apparent molecular weight of 96, 55 and 32 kDa, representing 17%, 27% and 24% of the total inhibin activity in the follicular fluid, respectively. The smallest inhibin, named 32 kDa bFF inhibin, that evidently suppressed the basal secretion of FSH from the pituitary cells, was purified to homogeneity with a 5330-fold purification factor in a recovery yield of ca. 11%. 32 kDa bFF inhibin thus purified consists of 2 polypeptide chains (A-chain: 20 kDa and B-chain: 13 kDa), linked by disulfide bridges. N-Terminal sequences were Ser-Thr-Pro-Pro- for the A-chain and Gly-Leu-Glu-Cys- for the B-chain. The identical N-terminal sequences were also found in 32 kDa pFF inhibin, except that Pro-3 of the bFF A-chain is substituted by Ala in the 20 kDa chain of pFF inhibin.

摘要

利用在蛋白质解离条件下进行的色谱法，对牛卵泡液（bFF）中约32 kDa的抑制素进行了纯化，这些条件是我们之前用于纯化猪卵泡液（pFF）抑制素时所建立的。在凝胶过滤中，bFF抑制素活性在3个不同区域洗脱，对应表观分子量分别为96、55和32 kDa，分别占卵泡液中总抑制素活性的17%、27%和24%。最小的抑制素，即32 kDa bFF抑制素，能明显抑制垂体细胞基础FSH分泌，经纯化达到同质，纯化因子为5330倍，回收率约为11%。如此纯化得到的32 kDa bFF抑制素由2条多肽链组成（A链：20 kDa，B链：13 kDa），通过二硫键相连。A链的N端序列为Ser-Thr-Pro-Pro-，B链的N端序列为Gly-Leu-Glu-Cys-。在32 kDa pFF抑制素中也发现了相同的N端序列，只是bFF A链的第3位Pro在pFF抑制素的20 kDa链中被Ala取代。