Kleinman H K, Ogle R C, Cannon F B, Little C D, Sweeney T M, Luckenbill-Edds L
Laboratory of Developmental Biology and Anomalies, National Institute of Dental Research, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1988 Feb;85(4):1282-6. doi: 10.1073/pnas.85.4.1282.
Laminin, a basement membrane glycoprotein promotes both cell attachment and neurite outgrowth. Separate domains on laminin elicit these responses, suggesting that distinct receptors occur on the surface of cells. NG108-15 neuroblastoma-glioma cells rapidly extend long processes in the presence of laminin. We report here that 125I-labeled laminin specifically binds to these cells and to three membrane proteins of 67, 110, and 180 kDa. These proteins were isolated by affinity chromatography on laminin-Sepharose. The 67-kDa protein reacted with antibody to the previously characterized receptor for cell attachment to laminin. Antibodies to the 110-kDa and 180-kDa bands demonstrated that the 110-kDa protein was found in a variety of epithelial cell lines and in brain, whereas the 180-kDa protein was neural specific. Antibodies prepared against the 110-kDa and 180-kDa proteins inhibited neurite outgrowth induced by the neurite-promoting domain of laminin, whereas antibodies to the 67-kDa laminin receptor had no effect on neurite outgrowth. We conclude that neuronal cells have multiple cell-surface laminin receptors and that the 110-kDa and 180-kDa proteins are involved in neurite formation.
层粘连蛋白是一种基底膜糖蛋白，可促进细胞黏附和神经突生长。层粘连蛋白上不同的结构域引发这些反应，这表明细胞表面存在不同的受体。在层粘连蛋白存在的情况下，NG108 - 15神经母细胞瘤-胶质瘤细胞会迅速伸出长长的突起。我们在此报告，125I标记的层粘连蛋白能特异性结合这些细胞以及三种分子量分别为67 kDa、110 kDa和180 kDa的膜蛋白。这些蛋白通过层粘连蛋白-琼脂糖亲和层析法分离得到。67 kDa的蛋白与针对先前已鉴定的层粘连蛋白细胞黏附受体的抗体发生反应。针对110 kDa和180 kDa条带的抗体表明，110 kDa的蛋白存在于多种上皮细胞系和脑组织中，而180 kDa的蛋白具有神经特异性。针对110 kDa和180 kDa蛋白制备的抗体抑制了由层粘连蛋白的神经突促进结构域诱导的神经突生长，而针对67 kDa层粘连蛋白受体的抗体对神经突生长没有影响。我们得出结论，神经元细胞具有多种细胞表面层粘连蛋白受体，并且110 kDa和180 kDa的蛋白参与神经突形成。
