Chemical control of metabolically-engineered voltage-gated K+ channels
Chemical control of metabolically-engineered voltage-gated K+ channels
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DOI：
10.1016/j.bmcl.2011.04.099
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发表时间：
2011-09-01
影响因子：
2.7
通讯作者：
Kobertz, William R.
中科院分区：
文献类型：
--
作者：
Hua, Zhengmao;Lvov, Anatoli;Kobertz, William R.
代谢寡糖工程是将具有独特功能基团的非天然聚糖安装到活细胞和动物的糖萼中的强有力的方法。使用这种方法，我们表明用含硫醇的唾液酸修饰的K+通道复合物被带有马来酰亚胺侧基的蝎子毒素不可逆地抑制。不可逆抑制需要糖基化的K+通道亚基，并且当连接毒素与马来酰亚胺的系链含有二硫键时，用温和的还原剂完全可逆。二硫键的切割不仅恢复了功能，而且将生物素部分递送到修饰的K+通道亚基，提供了优先标记在细胞中起作用的野生型K+通道复合物的新方法。(C)2011爱思唯尔有限公司保留所有权利。
Metabolic oligosaccharide engineering is a powerful approach for installing unnatural glycans with unique functional groups into the glycocalyx of living cells and animals. Using this approach, we showed that K+ channel complexes decorated with thiol-containing sialic acids were irreversibly inhibited with scorpion toxins bearing a pendant maleimide group. Irreversible inhibition required a glycosylated K+ channel subunit and was completely reversible with mild reductant when the tether connecting the toxin to the maleimide contained a disulfide bond. Cleavage of the disulfide bond not only restored function, but delivered a biotin moiety to the modified K+ channel subunit, providing a novel approach for preferentially labeling wild type K+ channel complexes functioning in cells. (C) 2011 Elsevier Ltd. All rights reserved.