Identification of a GM1-binding protein on the surface of murine neuroblastoma cells.
Identification of a GM1-binding protein on the surface of murine neuroblastoma cells.
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小鼠神经母细胞瘤细胞表面 GM1 结合蛋白的鉴定。
DOI：
10.1111/j.1471-4159.1992.tb09402.x
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发表时间：
1992
影响因子：
4.7
通讯作者：
Schengrund,CL
中科院分区：
文献类型：
--
作者：
Fueshko,SM;Schengrund,CL
S2OY小鼠神经母细胞瘤细胞似乎表达一种蛋白成分(S)，能够特异性地附着于GM1的寡糖部分(寡糖-GM1)。为了鉴定与寡聚GM1密切相关的蛋白质，制备了一种放射性标记的(125I)、可光激活的寡聚GM1的衍生物。这是通过将低聚甘油三酯的葡萄糖部分还原为1-脱氧-1-氨基葡萄糖醇，然后将胺与磺基琥珀酰亚胺2‐(p‐azidosalicylamido)ethyl‐1，3′‐dithiopropionate反应完成的。使用光活化探针的交联研究表明，它非常接近一种表观分子质量为∼71 kDa的蛋白质。在竞争实验中，由于∼71-kDa蛋白的摩尔过量为10倍，导致该蛋白的标记选择性减少；有必要与摩尔过量200倍的siayl乳糖预先孵育，以观察标记模式的相同变化，从而进一步支持了以下假设，即SS71-kDa蛋白与寡聚GM1特异性地结合。亚细胞分级和形态分析证实了寡聚GM1结合蛋白的细胞表面定位。
S2OY murine neuroblastoma cells appear to express a protein component(s) able to adhere specifically to the oligosaccharide portion of GM1 (oligo‐GM1). To identify proteins with which the oligo‐GM1 becomes closely associated, a radiolabeled (125I), photoactivatable derivative of oligo‐GM1 was prepared. This was accomplished by reductive amination of the glucosyl moiety of oligo‐GM1 to 1‐deoxy‐1‐aminoglucitol, followed by reaction of the amine with sulfosuccinimidyl 2‐(p‐azidosalicylamido)ethyl‐1,3′‐dithiopropionate (SASD). Crosslinking studies using the photoactivatable probe indicated that it came in close proximity to a protein with an apparent molecular mass of ∼ 71 kDa. In competition experiments, as little asa 10‐fold molar excess of oligo‐GM1 resulted in a selective reduction in labeling of this protein; preincubation with a 200‐fold molar excess of siayllactose was necessary to observe the same change in the labeling pattern, lending additional support to the hypothesis that the ∼ 71‐kDa protein specifically associates with oligo‐GM1. Cell surface location of the oligo‐GM1 binding protein was confirmed using subcellular fractionation and morphological analyses.