Dziadek M, Paulsson M, Timpl R
EMBO J. 1985 Oct;4(10):2513-8. doi: 10.1002/j.1460-2075.1985.tb03964.x.
Nidogen was purified in its genuine form with a mol. wt. of 150 000 (Nd-150) and as fragments with mol. wts. of 100 000 (Nd-100) and 80 000 (Nd-80) from a mouse tumor basement membrane by preventing activity of endogenous proteases with 6 M guanidine and protease inhibitors. The larger forms of nidogen were also identified in stable complexes with laminin in neutral salt extracts of the tumor and in cell culture medium. Purified Nd-150 and Nd-100, but not Nd-80, were shown to interact with laminin in various binding assays, albeit with lower potential than estimated for the genuine complexes formed in situ. Binding of Nd-150 and Nd-100 to fibronectin and to the globular domain of collagen IV was also observed, but not to heparan sulfate proteoglycan.
通过用6M胍和蛋白酶抑制剂抑制内源性蛋白酶的活性，从小鼠肿瘤基底膜中以其天然形式纯化了分子量为150000的巢蛋白（Nd - 150）以及分子量为100000（Nd - 100）和80000（Nd - 80）的片段。在肿瘤的中性盐提取物和细胞培养基中，还鉴定出巢蛋白的较大形式与层粘连蛋白形成稳定复合物。在各种结合试验中，纯化的Nd - 150和Nd - 100（而非Nd - 80）显示与层粘连蛋白相互作用，尽管其潜力低于原位形成的天然复合物的估计值。还观察到Nd - 150和Nd - 100与纤连蛋白以及IV型胶原的球状结构域结合，但不与硫酸乙酰肝素蛋白聚糖结合。
