VITRONECTIN - A MAJOR CELL ATTACHMENT-PROMOTING PROTEIN IN FETAL BOVINE SERUM
VITRONECTIN - A MAJOR CELL ATTACHMENT-PROMOTING PROTEIN IN FETAL BOVINE SERUM
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DOI：
10.1016/0014-4827(85)90173-9
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发表时间：
1985-01-01
影响因子：
3.7
通讯作者：
RUOSLAHTI, E
中科院分区：
文献类型：
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作者：
HAYMAN, EG;PIERSCHBACHER, MD;RUOSLAHTI, E
牛血清是大多数用于动物细胞培养的培养液的成分。血清的黏附促进作用一般归因于纤维连接蛋白，然而在牛血清中也经常有其他黏附促进分子的报道。利用一种通过SDS-PAGE分离粘附性蛋白的技术，我们图形化地证实了牛血清中存在第二种细胞附着蛋白，并提出了该分子相当于牛的Vitronectin的证据。这种蛋白质的分子大小与粘附性人血浆蛋白Vitronectin的大小相同。这种牛蛋白与人的Vitronectin也具有对玻璃的亲和力，可以通过玻璃微珠和离子交换层析相结合的方法进行纯化。分离的牛蛋白含有不同比例的80和65kD多肽。生长粘连蛋白B是一种血清多肽，其NH2末端序列与人玻璃连结蛋白完全相同。通过证明其NH2端氨基酸序列与人玻璃连蛋白和生长激素B有很强的同源性，从而确定其为玻璃连蛋白。对牛血浆和新鲜血清中纤维连接蛋白和玻璃连蛋白的黏附活性进行了定量测定，结果表明，玻璃连蛋白的结合活性高于纤维连接蛋白。在用于细胞培养的胎牛血清中，Vitronectin的优势尤为明显。在不同批次中，玻璃连蛋白的细胞黏附活性是纤维连接蛋白的8-16倍，使玻璃连蛋白成为常规细胞培养上清液中的主要黏附蛋白。
Bovine serum is a constituent of most media used for the culture of animal cells. The adhesion-promoting properties of serum are generally attributed to fibronectin, yet there have been frequent reports of other adhesion-promoting molecules in bovine serum. Using a technique in which adhesive proteins are visualized after separation by SDS-PAGE, we graphically confirm the presence of a second cell attachment protein in bovine serum and present the evidence that this molecule is the bovine equivalent of vitronectin. The molecular size of this protein is in the same range as the size of the adhesive human plasma protein, vitronectin. The bovine protein also shared with human vitronectin an affinity for glass, and it could be purified by a combination of glass bead and ion exchange chromatography. The isolated bovine protein had varying proportions of an 80 and a 65 kD polypeptide. It showed immunological cross-reactivity with anti-human vitronectin and with anti-human somatomedin B. Somatomedin B is a serum peptide which has a NH2-terminal sequence identical to that of human vitronectin. The identity of the bovine protein as vitronectin was established by showing that its NH2-terminal amino acid sequence is strongly homologous with those of human vitronectin and somatomedin B. Quantitation of the adhesive activities of fibronectin and vitronectin in bovine plasma and fresh serum showed that more activity is associated with vitronectin than with fibronectin. The preponderance of vitronectin was particularly clear in fetal bovine serum intended for cell culture. In various batches, cell attachment activity attributable to vitronectin was 8-16-fold greater than that of fibronectin, making vitronectin the main adhesive protein in routine cell culture media.