Risau W, Gautschi-Sova P, Böhlen P
Max-Planck-Institut für Entwicklungsbiologie, Tübingen, FRG.
EMBO J. 1988 Apr;7(4):959-62. doi: 10.1002/j.1460-2075.1988.tb02901.x.
We have investigated the nature of endothelial cell growth factors in 14-day embryonic and adult chick brain extracts. Mitogenic activity was isolated by a combination of cation-exchange, heparin-Sepharose affinity, and reverse-phase HPLC. Two major mitogenic fractions eluted from heparin-Sepharose at 0.8-1.3 M and 1.5-2 M. Biologically active proteins eluting at 0.8-1.3 M NaCl, after purification to homogeneity from embryonic and adult brain, were found to possess the same amino-terminal sequence as human acidic fibroblast growth factor (aFGF). The notion that the isolated mitogens represent chick aFGF is further supported by the findings that their affinity for heparin and their retention behavior in highly resolutive HPLC are indistinguishable from those of genuine aFGF. Mitogenic activities eluting at 1.5-2 M NaCl were also present in embryonic and adult brain, but in quantities insufficient for preliminary characterization. The high specific mitogenic activity for endothelial cells, high affinity for heparin and cross-reactivity with antibodies against bovine basic FGF (bFGF) suggest a relationship of those materials with basic FGF. Our data also suggest that the sequence of aFGF is highly conserved among vertebrates. While angiogenesis occurs predominantly in the embryonic brain, the absence of notable differences in the contents of the potent angiogenic factors aFGF and bFGF in embryonic versus adult chick brain is interesting.
我们研究了14天胚胎期和成年期鸡脑提取物中内皮细胞生长因子的性质。通过阳离子交换、肝素-琼脂糖亲和层析和反相高效液相色谱相结合的方法分离出促有丝分裂活性物质。从肝素-琼脂糖上洗脱下来的两个主要促有丝分裂组分分别在0.8 - 1.3M和1.5 - 2M处。从胚胎脑和成年脑纯化至同质后,在0.8 - 1.3M NaCl处洗脱的具有生物活性的蛋白质,其氨基末端序列与人酸性成纤维细胞生长因子(aFGF)相同。分离得到的促有丝分裂原代表鸡aFGF这一观点,进一步得到以下发现的支持:它们对肝素的亲和力以及在高分辨率高效液相色谱中的保留行为与真正的aFGF无法区分。在1.5 - 2M NaCl处洗脱的促有丝分裂活性物质在胚胎脑和成年脑中也存在,但数量不足以进行初步表征。这些物质对内皮细胞具有高特异性促有丝分裂活性、对肝素具有高亲和力且与抗牛碱性成纤维细胞生长因子(bFGF)抗体有交叉反应,表明它们与碱性成纤维细胞生长因子存在某种关系。我们的数据还表明,aFGF的序列在脊椎动物中高度保守。虽然血管生成主要发生在胚胎脑中,但胚胎期与成年期鸡脑相比,强效血管生成因子aFGF和bFGF的含量没有显著差异,这一点很有意思。