PURIFICATION AND CHARACTERIZATION OF GLIA MATURATION FACTOR-BETA - A GROWTH-REGULATOR FOR NEURONS AND GLIA
PURIFICATION AND CHARACTERIZATION OF GLIA MATURATION FACTOR-BETA - A GROWTH-REGULATOR FOR NEURONS AND GLIA
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DOI：
10.1073/pnas.86.10.3901
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发表时间：
1989-05-01
影响因子：
11.1
通讯作者：
ZAHEER, A
中科院分区：
文献类型：
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作者：
LIM, R;MILLER, JF;ZAHEER, A
通过改进纯化神经胶质成熟因子的程序，从牛脑中分离出一种蛋白质。该方法包括硫酸铵沉淀、DEAE-Sephacel、Sephadex G-75 和羟基磷灰石柱层析，通过肝素-Sepharose 柱，最后通过反相 HPLC 使用 C4 柱进行分级。分离的蛋白与小鼠单克隆抗体G2-09反应强烈，分子量为.apprxeq。 17,000，等电点 pH 4.9。 N 末端被封闭，但胰蛋白酶消化释放 28 个肽，其中 8 个已被测序。根据氨基酸组成估计，已知残基总数占整个 140 个残基蛋白质的三分之二以上，并且与任何已知蛋白质没有序列同源性。可逆的热复性大大增强了其生物活性。纯化的蛋白质刺激正常神经元和神经胶质细胞的分化。它抑制 N-18 神经母细胞瘤系和 C6 神经胶质瘤系的增殖，同时促进其表型表达。我们将这种蛋白质命名为神经胶质成熟因子.beta。
A protein has been isolated from bovine brains by using a modification of the procedure used to purify glia maturation factor. The method consists of ammonium sulfate precipitation, chromatography with DEAE-Sephacel, Sephadex G-75, and hydroxylapatite columns, passage through a heparin-Sepharose column, and finally fractionation by reverse-phase HPLC with a C4 column. The isolated protein reacts strongly with the mouse monoclonal antibody G2-09 and has a molecular weight of .apprxeq. 17,000 and an isoelectric point of pH 4.9. The N terminus is blocked, but tryptic digestion releases 28 peptides, 8 of which have been sequenced. The total known residues add up to more than two-thirds of the entire 140-residue protein, estimated from amino acid composition, and show no sequence homology with any known protein. Reversible thermal renaturation greatly enhances its biological activity. The purified protein stimulates differentiation of normal neurons as well as glial cells. It inhibits the proliferation of the N-18 neuroblastoma line and the C6 glioma line while promoting their phenotypic expression. We designate this protein glia maturation factor .beta.