Perderiset M, Courty J, Mereau A, Chevet E, Barritault D
Laboratoire de Recherche sur la Croissance Cellulaire, Université Paris Val-de-Marne, Créteil, France.
Biochimie. 1992 Dec;74(12):1091-6. doi: 10.1016/0300-9084(92)90007-2.
A new form of high affinity fibroblast growth factor receptor has been purified from adult bovine brain membranes. Purification was performed by chromatography on DEAE-Trisacryl and wheat germ agglutinin-agarose followed by FGF-2 affinity chromatography. Affinity labeling of purified fractions with 125I-FGF-2 showed after cross-linking a 170-kDa complex, suggesting the existence of a 150-kDa FGF receptor. No cross-reactivity with anti-FGF receptor 1 (FGFR-1 or flg) or with anti-receptor 2 (FGFR-2 or bek) antibodies could be detected with this partially purified receptor. Heparitinase treatment of the partially purified FGF receptor abolished the formation of the ligand receptor complex. The complex was restored in the presence of heparin in a dose dependent fashion, supporting the idea that heparin-like molecules are needed for proper binding. Further purification of the receptor was achieved by heparin-Sepharose affinity chromatography and yielded a purification of over 320,000-fold. The purified receptor fraction was radiolabeled and loaded on RPLC C4 column. Eluted fractions were analysed by SDS-PAGE. A major 150-kDa band was detected. These data show for the first time a new form of FGF receptor isolated from bovine brain membranes. This purified receptor displays affinity for heparin and was therefore named heparin binding FGF receptor (HB-FGFR). It remains unclear whether the receptor is a proteo-heparin sulfate or whether heparans are strongly associated and therefore are copurified. Large scale preparations are in progress for core protein structure studies.
一种新型高亲和力成纤维细胞生长因子受体已从成年牛脑膜中纯化出来。纯化过程通过在DEAE-三羟甲基氨基甲烷琼脂糖和麦胚凝集素琼脂糖上进行层析，随后进行FGF-2亲和层析。用125I-FGF-2对纯化组分进行亲和标记，交联后显示出一个170 kDa的复合物，表明存在一种150 kDa的FGF受体。用这种部分纯化的受体未检测到与抗FGF受体1（FGFR-1或flg）或抗受体2（FGFR-2或bek）抗体的交叉反应性。对部分纯化的FGF受体进行肝素酶处理可消除配体-受体复合物的形成。在肝素存在的情况下，复合物以剂量依赖的方式得以恢复，这支持了肝素样分子对于正确结合是必需的这一观点。通过肝素-琼脂糖亲和层析进一步纯化受体，纯化倍数超过320,000倍。将纯化的受体组分进行放射性标记并加载到反相高效液相色谱C4柱上。通过SDS-PAGE分析洗脱组分。检测到一条主要的150 kDa条带。这些数据首次表明从牛脑膜中分离出了一种新型的FGF受体。这种纯化的受体对肝素具有亲和力，因此被命名为肝素结合FGF受体（HB-FGFR）。目前尚不清楚该受体是蛋白聚糖硫酸酯还是硫酸乙酰肝素与之紧密结合并因此被共纯化。正在进行大规模制备以进行核心蛋白结构研究。
