Kalinin is more efficient than laminin in promoting adhesion of primary keratinocytes and some other epithelial cells and has a different requirement for integrin receptors.
Kalinin is more efficient than laminin in promoting adhesion of primary keratinocytes and some other epithelial cells and has a different requirement for integrin receptors.
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DOI：
10.1083/jcb.125.1.205
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发表时间：
1994-04
期刊：
影响因子：
--
通讯作者：
Aumailley M
中科院分区：
文献类型：
--
作者：
Rousselle P;Aumailley M
使用由mAb BM 165制备的免疫亲和柱，从鳞状细胞癌（SCC 25）用过的培养基中纯化Kalinin。亲和纯化的材料通过SDS-PAGE分离成165-155、140和105 kD的三条带，与从正常人角质形成细胞培养物中获得的那些相同，并且先前鉴定为激肽。Kalinin促进了大量正常细胞的粘附，并建立了具有类似于其他粘附分子如层粘连蛋白-巢蛋白复合物、纤连蛋白或胶原IV的活性的细胞系。然而，对于上皮来源的细胞（包括原代人角质形成细胞）的粘附，激肽是比层粘连蛋白-巢蛋白复合物好得多的底物。粘附到卡林蛋白后，细胞形状发生变化，从圆形细胞到完全展开的细胞，这取决于细胞类型。Kalinin的粘附促进活性是构象依赖性的，并通过热变性被废除。mAb BM 165阻止细胞粘附到kalinin，但不阻止细胞粘附到其他细胞外基质底物。然而，在不同的细胞中观察到完全或部分抑制，表明卡利宁分子上存在至少两个细胞结合位点。用功能阻断抗整联蛋白亚基抗体抑制细胞粘附的实验表明，α 3 β 1和α 6 β 1整联蛋白都参与与激肽的细胞相互作用，而对于与经典小鼠Engelbreth-Holm-Swarm层粘连蛋白的细胞粘附，只有α 6 β 1整联蛋白而不是α 3 β 1似乎是功能性的。总之，这些结果表明，激肽可能履行额外的功能比层粘连蛋白，特别是上皮细胞。
Kalinin was purified from squamous cell carcinoma (SCC25) spent culture media using an immunoaffinity column prepared from the mAb BM165. The affinity-purified material was separated by SDS-PAGE into three bands of 165-155, 140, and 105 kD identical to those obtained from normal human keratinocyte cultures and previously identified as kalinin. Kalinin promoted adhesion of a large number of normal cells and established cell lines with an activity similar to other adhesion molecules such as the laminin-nidogen complex, fibronectin, or collagen IV. However, kalinin was a much better substrate than laminin-nidogen complex for adhesion of cells of epithelial origin including primary human keratinocytes. Adhesion to kalinin was followed by cell shape changes ranging from rounded to fully spread cells depending on the cell types. The adhesion-promoting activity of kalinin was conformation dependent and was abolished by heat denaturation. mAb BM165 prevented cell adhesion to kalinin but not to other extracellular matrix substrates. However, either complete or partial inhibition was observed with different cells suggesting the existence of at least two cell- binding sites on the kalinin molecule. Experiments inhibiting cell adhesion with function-blocking anti-integrin subunit antibodies indicated that both alpha 3 beta 1 and alpha 6 beta 1 integrins are involved in the cellular interactions with kalinin, while for cell adhesion to classical mouse Engelbreth-Holm-Swarm laminin only alpha 6 beta 1 integrins, and not alpha 3 beta 1, appeared to be functional. Altogether, these results suggest that kalinin may fulfill additional functions than laminin, particularly for epithelial cells.