Metabolism of diazirine-modified N-acetylmannosamine analogues to photo-cross-linking sialosides.
Metabolism of diazirine-modified N-acetylmannosamine analogues to photo-cross-linking sialosides.
复制标题
DOI：
10.1021/bc2002117
复制
发表时间：
2011-09-21
影响因子：
4.7
通讯作者：
Kohler, Jennifer J.
中科院分区：
文献类型：
--
作者：
Bond, Michelle R.;Zhang, Haochi;Kim, Jaekuk;Yu, Seok-Ho;Yang, Fan;Patrie, Steven M.;Kohler, Jennifer J.
末端唾液酸残基通常介导细胞表面糖缀合物的相互作用。唾液酸依赖性相互作用通常表现出快速的解离速率，排除了使用传统的生物技术进行复杂的分离。为了稳定这些短暂的相互作用，我们采用了有针对性的光交联方法，其中通过使用代谢寡糖工程将二氮丙啶光交联剂掺入细胞表面唾液酸化的糖缀合物中。我们描述了三个双吖丙啶修饰的N-乙酰甘露糖胺（ManNAc）类似物，其中吡喃糖环和双吖丙啶之间的接头的长度是不同的。这些类似物各自代谢成其各自的唾液酸对应物，其被添加到糖蛋白和糖脂两者中。二氮丙啶修饰的唾液酸类似物可以掺入α2-3和α2-6键中。暴露于UV照射后，二氮丙啶修饰的糖缀合物共价交联到它们的相互作用伴侣。我们证明，所有三个二氮丙啶修饰的类似物能够与内源性唾液酸竞争，尽管程度不同。我们发现，较大的类似物代谢效率较低，但仍然可以作为有效的交联剂。值得注意的是，添加二氮杂环丙烯取代基会干扰ManNAc类似物代谢成除唾液酸糖苷以外的聚糖，从而提供选择性地将交联剂掺入唾液酸化分子中的保真度。这些化合物是无毒的，在交联研究所需的浓度下仅显示出最小的生长抑制。本报告提供了必要的信息部署的光交联类似物捕捉和研究短暂的，但必不可少的，唾液酸介导的相互作用。
Terminal sialic acid residues often mediate the interactions of cell surface glycoconjugates. Sialic acid-dependent interactions typically exhibit rapid dissociation rates, precluding the use of traditional biological techniques for complex isolation. To stabilize these transient interactions, we employ a targeted photocrosslinking approach in which a diazirine photocrosslinker is incorporated into cell surface sialylated glycoconjugates through the use of metabolic oligosaccharide engineering. We describe three diazirine-modified N-acetylmannosamine (ManNAc) analogs in which the length of the linker between the pyranose ring and the diazirine was varied. These analogs were each metabolized to their respective sialic acid counterparts, which were added to both glycoproteins and glycolipids. Diazirine-modified sialic acid analogs could be incorporated into both α2–3 and α2–6 linkages. Upon exposure to UV irradiation, diazirine-modified glycoconjugates were covalently crosslinked to their interaction partners. We demonstrate that all three diazirine-modified analogs were capable of competing with endogeneous sialic acid, albeit to varying degrees. We found that larger analogs were less efficiently metabolized, yet could still function as effective crosslinkers. Notably, the addition of the diazirine substituent interferes with metabolism of ManNAc analogs to glycans other than sialosides, providing fidelity to selectively incorporate the crosslinker into sialylated molecules. These compounds are non-toxic and display only minimal growth inhibition at the concentrations required for crosslinking studies. This report provides essential information for the deployment of photocrosslinking analogs to capture and study ephemeral, yet essential, sialic acid-mediated interactions.
登录
查看更多内容
影响因子：
6.5
作者：
Bussink, Anton P.;van Swieten, Paul F.;Overkleeft, Herman S.
通讯作者：
Overkleeft, Herman S.
DOI：
10.1111/j.1432-1033.1982.tb19760.x
发表时间：
1982-01-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
IGLESIAS, JL;LIS, H;SHARON, N
通讯作者：
SHARON, N
影响因子：
4.8
作者：
Luchansky, SJ;Yarema, KJ;Bertozzi, CR
通讯作者：
Bertozzi, CR
DOI：
10.1006/bbrc.1998.9650
发表时间：
1998-12-18
影响因子：
3.1
作者：
Keppler, OT;Herrmann, M;Pawlita, M
通讯作者：
Pawlita, M
影响因子：
2.9
作者：
LADISCH, S;GILLARD, B
通讯作者：
GILLARD, B