Synthesis and Evaluation of a Photoactive Probe with a Multivalent Carbohydrate for Capturing Carbohydrate鈥揕ectin Interactions

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• 作者：Tsung-Che Chang ; Chian-Hui Lai ; Chih-Wei Chien ; Chien-Fu Liang ; Avijit Kumar Adak ; Yung-Jen Chuang ; Yu-Ju Chen ; Chun-Cheng Lin
• 刊名：Bioconjugate Chemistry
• 出版年：2013
• 出版时间：November 20, 2013
• 年：2013
• 卷：24
• 期：11
• 页码：1895-1906
• 全文大小：501K
• 年卷期：v.24,no.11(November 20, 2013)
• ISSN：1520-4812

文摘

Lectins are ubiquitous carbohydrate-binding proteins of nonimmune origin that are characterized by their specific recognition of defined monosaccharide or oligosaccharide structures. However, the use of carbohydrates to study lectin has been restricted by the weak binding affinity and noncovalent character of the interaction between carbohydrates and lectin. In this report, we designed and synthesized a multifunctional photoaffinity reagent composed of a trialkyne chain, a masked latent amine group, and a photoreactive 3-trifluoromethyl-3-phenyl-diazirine group in high overall yield. Two well-defined chemistries, Huisgen-Sharpless click chemistry and amide bond coupling, were the key steps for installing the multivalent character and tag in our designed photoaffinity probe. The photolabeling results demonstrated that the designed probe selectively labeled the target lectin, RCA₁₂₀ (Ricinus communis Agglutinin), in an E. coli lysate and an asialoglycoprotein receptor (ASGP-R) on intact HepG2 cell membranes. Moreover, the probe also enabled the detection of weak protein鈥損rotein interactions between RCA₁₂₀ and ovalbumin (OVA).