化学糖生物学教学课件金属离子探针.ppt

So, with all those introductions. Here comes our experimental setup. We simply inject the azidosugars into rats, and run the reaction inside the animals, and then image the hearts undermicroscope. My student gives this project a nice name: sweet heart, because we are studying sugars in the hearts. * Iso: isopro terenol Furthermore, we used our imaging strategy to look at the pathological process, hypertrophy, which is the enlargement of the heart and often leads to heart failure. We found the O-linked glycosyaltion was upregulated during hypertrophy. The underline mechanism is under investegation. * * * * 1. Relatively low level of drug incorporation, up to 2 molecules per IgG 2. Relies on the presence of core fucose moiety 3. May inhibit antibody effector functions * enrichment with streptavidin beads Proteomic analysis of tumor sialylglycoproteins m/z LC-MS/MS Proteomic analysis of tumor sialylglycoproteins Day 14 Day 18 Day 21 Day 14 Day 18 Day 21 X. Chen, et al. Angew. Chem. Int. Ed. DOI: 10.1002/anie.201408442 A FRET-based method for protein-specific glycan imaging Donor Acceptor FRET：F?rster Resonance Energy Transfer FRET Challenges: thousands of glycoproteins share same glycans dynamic glycosylation X. Chen, et al. J. Am. Chem. Soc. 136, 679 (2014). Dual labeling of a specific receptor Direct Raman imaging of glycosylation Chemical reporter strategy: Two step process Hinges on bioorthogonal reactions Fluorophores are huge Fluorescent Raman reporter strategy: One step process Reaction-free labeling Better reporter Surface enhanced Raman scattering (SERS) detection SERS detection of SiaNAl on cell surfaces X. Chen, Z. Tian, et al. Angew. Chem. Int. Ed. 52, 7266 (2013). “Sweet Heart”: Visualizing cardiac glycome in intact hearts Metabolic labeling for 7 days N3 N3 DBCO-Fluor 488 Copper-free click chemistry Isolate heart Sialylation is upregulated in hypertrophy X. Chen, et al. J. Am. Chem. Soc. DOI: 10.1021/ja508484c 用化学-酶标记策略分离鉴定O-GlcNAc