SWAINSONINE - AN INHIBITOR OF GLYCOPROTEIN PROCESSING
SWAINSONINE - AN INHIBITOR OF GLYCOPROTEIN PROCESSING
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DOI：
10.1073/pnas.78.12.7393
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发表时间：
1981-01-01
期刊：
影响因子：
--
通讯作者：
VOSBECK, K
中科院分区：
文献类型：
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作者：
ELBEIN, AD;SOLF, R;VOSBECK, K
Swainsonine 是一种吲哚里西啶生物碱（来自 Swainsona canescens），可抑制无细胞提取物和培养的动物细胞中天冬酰胺衬里的糖蛋白的加工。在肝脏颗粒酶制剂中，0.1-1.0μM的苦马豆素抑制从高甘露糖糖肽释放[3H]甘露糖的甘露糖苷酶，但仅轻微抑制从葡萄糖标记的糖肽释放葡萄糖。培养中的 MDCK [犬肾上皮] 和中国仓鼠卵巢细胞将 [2-3H] 甘露糖和 [6-3H]-葡萄糖胺掺入高甘露糖和复合类型的寡糖中。当这些细胞与八氢吲嗪二醇一起孵育，然后用甘露糖或葡萄糖胺标记时，复合型糖肽中的标记量显着减少，而高甘露糖型中的放射性显着增加。这种变化是通过放射性的增加来监测的，随着苦马苷浓度的增加，放射性变得容易被内切氨基葡萄糖苷酶 H 消化。苦马豆素处理的细胞中的内切氨基葡萄糖苷酶 H 释放的寡糖比对照细胞更大且更均匀，并在 Man9GlcNAc 位置从 Bio-Gel P-4 中洗脱。几种组织培养细胞系在苦马豆素存在下生长，以确定其对细胞表面糖蛋白的影响。在生物碱中生长的细胞显示出结合大肠杆菌 B886（一种与高甘露糖糖蛋白结合的细菌）的能力增强。这些细胞还表现出 [3H]伴刀豆球蛋白 A 的结合增加。
Swainsonine, an indolizidine alkaloid [from Swainsona canescens], inhibits the processing of asparagine-lined glycoproteins in both cell-free extracts and animal cells in culture. In liver particulate enzyme preparation, swainsonine at 0.1-1.0 .mu.M inhibited the mannosidase that releases [3H]mannose from a high mannose glycopeptide but only slightly inhibited the release of glucose from a glucose-labeled glycopeptide. MDCK [canine kidney epithelium] and Chinese hamster ovary cells in culture incorporate [2-3H]mannose and [6-3H]-glucosamine into both high mannose and complex types of oligosaccharides. When these cells were incubated with swainsonine and then labeled ith mannose or glucosamine, there was a dramatic decrease in the amount of label in the complex type of glycopeptide and a substantial increase in the radioactivity in the high mannose type. This change was monitored by the increase in radioactivity that became susceptible to digestion by endoglucosaminidase H with increasing concentrations of swainosine. The endoglucosaminidase H-released oligosaccharide(s) from swainsonine-treated cells was larger and more homogenous than that from controls and eluted from Bio-Gel P-4 at the position of Man9GlcNAc. Several tissue culture cell lines were grown in the presence of swainsonine to determine its effect on cell surface glycoproteins. Cells grown in the alkaloid showed an increased capacity to bind Escherichia coli B886, a bacterium that binds to high mannose glycoproteins. These cells also showed an increasing binding of [3H]concanavalin A.