LG4-5 domains of laminin-211 binds alpha-dystroglycan to allow myotube attachment and prevent anoikis.
LG4-5 domains of laminin-211 binds alpha-dystroglycan to allow myotube attachment and prevent anoikis.
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laminin-211 的 LG4-5 结构域结合 α-肌营养不良聚糖，以允许肌管附着并防止失巢凋亡。
DOI：
10.1002/jcp.21927
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发表时间：
2010
影响因子：
5.6
通讯作者：
Jarrett,HarryW
中科院分区：
文献类型：
--
作者：
Munoz,Jesus;Zhou,Yanwen;Jarrett,HarryW
聚(甲基丙烯酸羟乙酯)(PolyHEMA)防止细胞附着在小鼠C2C12肌管中用于研究失巢凋亡，这是一种细胞在未附着或附着到不适当的基质时死亡的过程。建立了一种有效地将蛋白质包埋到聚HEMA中的方法，并测定了其对培养的肌管的影响。生长在PolyHEMA涂层平板上的肌管不能附着在表面并以圆形悬浮细胞的形式存在，细胞凋亡和坏死急剧增加，活细胞数量减少。掺入层粘连蛋白(Laminin-211)或层粘连蛋白-α2链C末端的短层粘连蛋白球状(LG4-5)模块(称为2E3)可以减少α-营养不良糖链的凋亡和坏死并提高存活率，而牛血清白蛋白的作用要小得多，显示了这种作用对这些基质蛋白的特异性。一种结合层粘连蛋白的肌膜受体是α-DystroGan。与α结合但不阻断层粘连蛋白结合的抗体(VIA4)对Merosin或2E3包埋平板的凋亡或存活率影响不大，而另一种特异性阻断结合的抗体(IIH6)则显着降低存活率并增加细胞凋亡率。当将Merosin或2E3添加到培养基中而不是包埋在平板上时，即使细胞仍处于悬浮状态，这些也可以提高存活率和减少凋亡，尽管效果不如细胞附着的嵌入蛋白那么大。因此，我们得出结论，层粘连蛋白211的小LG4-5模块与α-营养不良多糖的结合在防止失巢失巢方面是重要的，而附着加结合是最大限度地保持细胞存活所必需的。J.细胞。物理。222：111-119,2010。©2009威利-利斯，Inc.
Poly(2‐hydroxyethyl methacrylate) (PolyHEMA) prevents cell attachment was used here to study anoikis, the process where cells die when unattached or attached to an inappropriate matrix, in mouse C2C12myotubes. A method was developed to efficiently embed proteins into PolyHEMA and the effect on cultured myotubes was determined. Myotubes grown on PolyHEMA‐coated plates fail to attach to the surface and remain as rounded, suspended cells, undergo dramatic increases in apoptosis and necrosis, and the number of viable cells decreases. Incorporation of merosin (laminin‐211) or the short laminin globular (LG4–5) modules of the laminin‐α2 chain C‐terminus (called 2E3) that binds α‐dystroglycan diminishes both apoptosis and necrosis and increases viability while bovine serum albumin had a much lesser effect, showing the specificity of this effect for these matrix proteins. One sarcolemma receptor for laminin‐binding is α‐dystroglycan. An antibody which binds α‐dystroglycan but which does not block laminin‐binding (VIA4) had little effect on apoptosis or viability on merosin or 2E3 embedded plates while another antibody (IIH6) which specifically blocks binding dramatically decreased viability and increased apoptosis. When merosin or 2E3 are added to culture media rather than embedded on plates these can also increase viability and decrease apoptosis even though the cells remain in suspension, though the effect is not as great as found for the embedded proteins where the cells attach. Thus, we conclude that the binding of a small LG4–5 modules of laminin‐211 to α‐dystroglycan is important in preventing anoikis and that attachment plus binding is necessary for maximal cell survival. J. Cell. Physiol. 222:111–119, 2010. © 2009 Wiley‐Liss, Inc.