HUMAN TRANSFORMING GROWTH-FACTOR TYPE-BETA-2 - PRODUCTION BY A PROSTATIC ADENOCARCINOMA CELL-LINE, PURIFICATION, AND INITIAL CHARACTERIZATION
HUMAN TRANSFORMING GROWTH-FACTOR TYPE-BETA-2 - PRODUCTION BY A PROSTATIC ADENOCARCINOMA CELL-LINE, PURIFICATION, AND INITIAL CHARACTERIZATION
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DOI：
10.1021/bi00383a002
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发表时间：
1987-05-05
期刊：
影响因子：
2.9
通讯作者：
MARQUARDT, H
中科院分区：
文献类型：
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作者：
IKEDA, T;LIOUBIN, MN;MARQUARDT, H
人β2转化生长因子(hTGFβ2)是从人前列腺癌PC-3细胞株培养的添加他莫昔芬的无血清培养基中纯化的。HTGF-β2的纯化通过生长抑制实验进行监测，并通过甲基硅基控制的孔玻璃批量纯化，然后是凝胶渗透层析和反相高效液相色谱。人转化生长因子-β2的总回收率为初始活性的75%，并产生22µg的人转化生长因子-β2/L。在含10%胎牛血清的条件下，人转化生长因子-β2对MV1Lu水貂肺上皮细胞(CCL-)的半最大抑制作用所需的浓度约为下午5点。纯化的hTGF-β2经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法分析，其相对分子质量为24,000，由两条二硫键连接的明显相同的多肽链组成，相对分子质量为13,000。测定了人转化生长因子β2的氨基末端序列。人转化生长因子-β2和转化生长因子-β的氨基酸序列比对。揭示了具有统计学意义的序列同源性。基于广泛的氨基酸序列同源性，我们建议将这种新分离的多肽命名为转化生长因子-β2。报道的结果表明，转化生长因子-β。(转化生长因子-β1)和转化生长因子-β2可能是从一个共同的前体进化而来的。
Human type .beta.2 transforming growth factor (hTGF-.beta.2) was purified from tamoxifen-supplemented, serum-free medium conditioned by the human prostatic adenocarcinoma cell line PC-3. The purification of hTGF-.beta.2 was monitored in a growth inhibition assay and was achieved by batch purification on methylsilyl-controlled pore glass, followed by gel permeation chromatography and reversed-phase high-performance liquid chromatography. The overall recovery of hTGF-.beta.2 was 75% of the initial activity and yielded 22 .mu.g of hTGF-.beta.2/L of conditioned medium. The concentration of hTGF-.beta.2 required for half-maximal inhibition of Mv 1 Lu mink lung epithelial cells (CCl-64) was approximately 5 pM when assayed in the presence of 10% fetal bovine serum. The purified hTGF-.beta.2 has a molecular weight of 24,000 when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and consists of two disulfide-linked, apparently identical polypeptide chains, with a molecular weight of 13,000. The amino-terminal sequence of hTGF-.beta.2 was determined. Alignment of the amino acid sequences of hTGF-.beta.2 and hTGF-.beta. reveals statistically significant sequence homology. On the basis of the extensive amino acid sequence homology, we propose the term TGF-.beta.2 for this newly isolated polypeptide. The reported results suggest that TGF-.beta. (TGF-.beta.1) and TGF-.beta.2 may have evolved from a common progenitor.