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视网膜来源的眼源性生长因子的纯化、特性及生物学特性:与脑源性生长因子的类比

Purification, characterization, and biological properties of the eye-derived growth factor from retina: analogies with brain-derived growth factor.

作者信息

Barritault D, Plouët J, Courty J, Courtois Y

出版信息

J Neurosci Res. 1982;8(2-3):477-90. doi: 10.1002/jnr.490080235.

Abstract

Several ocular tissues contain a polypeptide growth factor(s) (eye-derived growth factor(s) or EDGF) that stimulates the proliferation of a large variety of cells from different tissues or species. Partial purification of EDGF from adult bovine retina was accomplished by acid precipitation, blue Ultrogel chromatography, and high-pressure liquid chromatography. EDGF has an isoelectrical point at pH 4.5 +/- 0.5 and an apparent molecular weight of 17,500 +/- 3,500 measured by high-pressure liquid chromatography. On the basis of these data as well as its biological properties EDGF is different from other known growth factors. The specific activity of highly purified EDGF is 1,000-fold greater than that of a 20,000g supernatant of a crude retinal extract. At this stage of purification, EDGF stimulates replicative DNA synthesis and cell proliferation of bovine epithelium lens cells at a concentration of 14 ng/ml of culture medium and for these cells is as efficient as purified brain fibroblast growth factor. The same purification steps were applied to crude bovine brain extracts. Growth factor activity was recovered exactly as for EDGF with slightly smaller apparent molecular weight 14,000 +/- 3,500, suggesting a great similarity between the two tissues as a source of growth factors. A purification of about 2,500-fold was obtained and cell proliferation stimulated at a concentration of 100 ng/ml. Interestingly an inhibitory activity not retained on blue Ultrogel was recovered from both preparations.

摘要

几种眼组织含有一种多肽生长因子(眼源性生长因子或EDGF),它能刺激来自不同组织或物种的多种细胞增殖。通过酸沉淀、蓝色琼脂糖凝胶色谱法和高压液相色谱法从成年牛视网膜中部分纯化了EDGF。EDGF的等电点在pH 4.5±0.5,通过高压液相色谱法测得其表观分子量为17,500±3,500。基于这些数据及其生物学特性,EDGF与其他已知生长因子不同。高度纯化的EDGF的比活性比粗视网膜提取物20,000g上清液的比活性高1000倍。在纯化的这个阶段,EDGF在培养基浓度为14 ng/ml时刺激牛晶状体上皮细胞的复制性DNA合成和细胞增殖,并且对于这些细胞而言,其效率与纯化的脑成纤维细胞生长因子相同。相同的纯化步骤应用于粗牛脑提取物。生长因子活性的回收情况与EDGF完全相同,表观分子量略小,为14,000±3,500,这表明作为生长因子来源的这两种组织之间有很大的相似性。获得了约2500倍的纯化倍数,在浓度为100 ng/ml时刺激细胞增殖。有趣的是,从这两种制剂中都回收了未保留在蓝色琼脂糖凝胶上的抑制活性。

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