Institute of Biophysics, The Czech Academy of Sciences, Královopolská 135, 612 65, Brno, Czech Republic; Department of Biochemistry, Faculty of Science, Masaryk University, Kotlářská 2, 611 37, Brno, Czech Republic.
Institute of Biophysics, The Czech Academy of Sciences, Královopolská 135, 612 65, Brno, Czech Republic.
Bioelectrochemistry. 2023 Aug;152:108457. doi: 10.1016/j.bioelechem.2023.108457. Epub 2023 May 10.
Trisaccharides bind to their interaction partners-lectins relatively weakly, which makes detection of their complexes challenging. In this work, we show that an osmolyte presence improves the distinguishing complexes of lectin Sambucus nigra with trisialyllactoses with various binding affinities. The addition of osmolyte, non-binding sugar mannose significantly improved the precision of binding experiments performed using chronopotentiometric stripping at the electrode surface and fluorescence analysis in solution. Osmolytes minimized nonspecific interactions between binding sugar and lectin. Obtained findings can be utilized in any in vitro methods studying interactions of carbohydrates, respectively their conjugates with proteins. The study of carbohydrate interactions appears important since they play essential roles in a variety of biological processes including carcinogenesis.
三糖与它们的相互作用伙伴——凝集素的结合相对较弱，这使得检测它们的复合物具有挑战性。在这项工作中，我们表明，渗透剂的存在改善了凝集素 Sambucus nigra 与具有不同结合亲和力的各种三唾液酸乳糖的复合物的区分。添加渗透剂、非结合糖甘露糖显著提高了在电极表面使用恒电流溶出伏安法和溶液荧光分析进行结合实验的精度。渗透剂最小化了结合糖和凝集素之间的非特异性相互作用。所获得的发现可用于研究碳水化合物及其与蛋白质的缀合物相互作用的任何体外方法。研究碳水化合物的相互作用很重要，因为它们在包括致癌作用在内的各种生物过程中起着重要作用。
