Glycan Imaging in Intact Rat Hearts and Glycoproteomic Analysis Reveal the Upregulation of Sialylation during Cardiac Hypertrophy
Glycan Imaging in Intact Rat Hearts and Glycoproteomic Analysis Reveal the Upregulation of Sialylation during Cardiac Hypertrophy
复制标题
完整大鼠心脏的聚糖成像和糖蛋白组学分析揭示了心脏肥大期间唾液酸化的上调。
DOI：
10.1021/ja508484c
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发表时间：
2014-12-17
影响因子：
15
通讯作者：
Chen, Xing
中科院分区：
文献类型：
--
作者：
Rong, Jie;Han, Jing;Chen, Xing
在心脏中，糖基化参与多种生理和病理过程。心脏糖基化是动态调节的，其在体内监测仍然具有挑战性。在这里，我们描述了一种化学方法来分析动态心脏糖代谢标记的心脏聚糖与叠氮糖在活的大鼠。叠氮化物，作为一个化学报告，化学选择性共轭与荧光团使用无铜点击化学聚糖成像;衍生叠氮化物与亲和标签允许富集和糖基化的心脏蛋白的蛋白质组学鉴定。我们通过观察完整心脏中的心脏唾液酸化聚糖和鉴定200多种用唾液酸修饰的心脏蛋白来证明这种方法。我们进一步应用这种方法来研究肥大心脏中的唾液酸化。影像学结果显示，诱导心肌肥大后唾液酸生物合成增加。定量蛋白质组学分析鉴定了多种唾液酸化蛋白，包括在肥大过程中上调的神经细胞粘附分子1、T-激肽原和α 2-巨球蛋白。该方法可以进一步扩展到其他类型的糖基化，如粘蛋白型0-连接的糖基化所例示的。我们的研究结果突出了代谢聚糖标记结合生物正交化学在探索心脏糖组的生物合成和功能在病理生理反应中的应用。
In the heart, glycosylation is involved in a variety of physiological and pathological processes. Cardiac glycosylation is dynamically regulated, which remains challenging to monitor in vivo. Here we describe a chemical approach for analyzing the dynamic cardiac glycome by metabolically labeling the cardiac glycans with azidosugars in living rats. The azides, serving as a chemical reporter, are chemoselectively conjugated with fluorophores using copper-free click chemistry for glycan imaging; derivatizing azides with affinity tags allows enrichment and proteomic identification of glycosylated cardiac proteins. We demonstrated this methodology by visualization of the cardiac sialylated glycans in intact hearts and identification of more than 200 cardiac proteins modified with sialic acids. We further applied this methodology to investigate the sialylation in hypertrophic hearts. The imaging results revealed an increase of sialic acid biosynthesis upon the induction of cardiac hypertrophy. Quantitative proteomic analysis identified multiple sialylated proteins including neural cell adhesion molecule 1, T-kininogens, and alpha(2)-macroglobulin that were upregulated during hypertrophy. The methodology may be further extended to other types of glycosylation, as exemplified by the mucin-type O-linked glycosylation. Our results highlight the applications of metabolic glycan labeling coupled with bioorthogonal chemistry in probing the biosynthesis and function of cardiac glycome during pathophysiological responses.