Isolation and biological properties of osteopontin from bovine milk
Isolation and biological properties of osteopontin from bovine milk
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DOI：
10.1006/prep.1996.0699
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发表时间：
1997-04-01
影响因子：
1.6
通讯作者：
Meininger, GA
中科院分区：
文献类型：
--
作者：
Bayless, KJ;Davis, GE;Meininger, GA
采用离子交换和疏水层析法从牛乳中分离骨桥蛋白。DEAE-Sephacel柱，然后双苯基-Sepharose柱产生类似于每升牛奶8 mg的纯化蛋白。SDS-PAGE分析显示蛋白质在M（r）60，000处迁移。前7个氨基酸的NH 2-末端序列分析显示该蛋白与先前报道的牛OPN相同。此外，我们的制备证明了OPN的预期生物学特性，包括内皮细胞和血管平滑肌细胞以剂量依赖性和Arg-Gly-Asp依赖性方式粘附于OPN。此外，偶联到琼脂糖凝胶的OPN能够结合来自内皮细胞去污剂提取物的α（v）β（3）整联蛋白。因此，我们的方法从丰富的天然来源中产生了具有生物活性的OPN。(C)北京：科学出版社.
A procedure for the isolation of osteopontin (OPN) from bovine milk using ion-exchange and hydrophobic chromatography is described. A DEAE-Sephacel column followed by dual phenyl-Sepharose columns yielded similar to 8 mg of purified protein per liter of milk. SDS-PAGE analysis revealed that the protein migrated at M(r) 60,000. NH2-terminal sequence analysis of the first seven amino acids revealed the protein to be identical to that previously reported for bovine OPN. Also, our preparation demonstrated expected biological properties of OPN including adhesion of both endothelial and vascular smooth muscle cells to OPN in a dose- and Arg-Gly-Asp dependent manner. Furthermore, OPN coupled to Sepharose was capable of binding the alpha(v) beta(3) integrin from a detergent extract of endothelial cells. Thus, our procedure yielded biologically active OPN from an abundant and natural source. (C) 1997 Academic Press.