Akgun Burcin, Hall Dennis G
Department of Chemistry, 4-010 CCIS, University of Alberta, Edmonton, Alberta, T6G 2G2, Canada.
Angew Chem Int Ed Engl. 2016 Mar 14;55(12):3909-13. doi: 10.1002/anie.201510321. Epub 2016 Feb 23.
A new click bioorthogonal reaction system was devised to enable the fast ligation (k(ON) ≈340 m(-1) s(-1)) of conjugatable derivatives of a rigid cyclic diol (nopoldiol) and a carefully optimized boronic acid partner, 2-methyl-5-carboxymethylphenylboronic acid. Using NMR and fluorescence spectroscopy studies, the corresponding boronates were found to form reversibly within minutes at low micromolar concentration in water, providing submicromolar equilibrium constant (K(eq) ≈10(5) -10(6) m(-1)). Efficient protein conjugation under physiological conditions was demonstrated with model proteins thioredoxin and albumin, and characterized by mass spectrometry and gel electrophoresis.
设计了一种新的点击生物正交反应体系，以实现刚性环状二醇（诺波二醇）的可共轭衍生物与精心优化的硼酸伴侣2-甲基-5-羧甲基苯硼酸的快速连接（k(ON)≈340 m⁻¹ s⁻¹）。通过核磁共振和荧光光谱研究发现，相应的硼酸酯在水中低微摩尔浓度下几分钟内即可可逆形成，提供亚微摩尔级的平衡常数（K(eq)≈10⁵ - 10⁶ m⁻¹）。用模型蛋白硫氧还蛋白和白蛋白证明了在生理条件下的高效蛋白质共轭，并通过质谱和凝胶电泳进行了表征。
