Center for Cancer and Developmental Biology, California State University, Northridge, 18111 Nordhoff Street, Northridge, CA 91330-8303, USA.
Exp Cell Res. 2010 Aug 1;316(13):2204-11. doi: 10.1016/j.yexcr.2010.04.026. Epub 2010 May 8.
We present an unusual and novel model for initial investigations of a putative role for specifically conformed glycans in cellular interactions. We have used alpha- and ss-amylase and alpha- and ss-glucosidase in dose-response experiments evaluating their effects on archenteron organization using the NIH designated sea urchin embryo model. In quantitative dose-response experiments, we show that defined activity levels of alpha-glucosidase and ss-amylase inhibited archenteron organization in living Lytechinus pictus gastrula embryos, whereas all concentrations of ss-glucosidase and alpha-amylase were without substantial effects on development. Product inhibition studies suggested that the enzymes were acting by their specific glycosidase activities and polyacrylamide gel electrophoresis suggested that there was no detectable protease contamination in the active enzyme samples. The results provide evidence for a role of glycans in sea urchin embryo cellular interactions with special reference to the possible structural conformation of these glycans based on the differential activities of the alpha- and ss-glycosidases.
我们提出了一个不寻常的新模型,用于初步研究特定构象聚糖在细胞相互作用中可能发挥的作用。我们使用α-和 ss-淀粉酶以及α-和 ss-葡萄糖苷酶进行了剂量反应实验,使用 NIH 指定的海胆胚胎模型评估了它们对原肠胚组织的影响。在定量剂量反应实验中,我们表明,定义的α-葡萄糖苷酶和 ss-淀粉酶活性水平抑制了活体 Lytechinus pictus 原肠胚胚胎的原肠胚组织,而 ss-葡萄糖苷酶和α-淀粉酶的所有浓度对发育均无明显影响。产物抑制研究表明,这些酶是通过其特定的糖苷酶活性起作用的,聚丙烯酰胺凝胶电泳表明在活性酶样品中没有检测到蛋白酶污染。这些结果为糖在海胆胚胎细胞相互作用中的作用提供了证据,特别是基于α-和 ss-糖苷酶的不同活性,对这些糖的可能结构构象进行了特别参考。