Bisaryl hydrazones as exchangeable biocompatible linkers.
Bisaryl hydrazones as exchangeable biocompatible linkers.
复制标题
DOI：
10.1002/anie.200906756
复制
发表时间：
2010-03-08
影响因子：
16.6
通讯作者：
Dawson, Philip E.
中科院分区：
文献类型：
--
作者：
Dirksen, Anouk;Yegneswaran, Subramanian;Dawson, Philip E.
从复杂的生物混合物中选择性富集标记的分子对于化学生物学和蛋白质组学中的方法是至关重要的。使用生物素作为亲和标签在（strept）抗生物素蛋白珠上进行亲和纯化是实现这一目标的最广泛应用的方法之一，充分利用了生物素对（strept）抗生物素蛋白的高亲和力（Ka = 1.7× 1015 M− 1）[1]。然而，该方法受到洗脱所需的苛刻变性条件的限制，例如SDS煮沸或用8 M胍（pH 1.5）处理。蛋白质结构和功能丧失，靶蛋白可能被非特异性结合到珠上的蛋白质污染。已经探索了两种策略来在温和条件下进行酶解：i）通过调节Ka来减弱生物素-（链霉）抗生物素蛋白相互作用[2]和ii）引入蛋白水解[3]或化学[4]可切割的接头。虽然第一种策略确实改善了生物素化蛋白从（链霉菌）抗生物素蛋白珠粒的释放，但它对固定的严格性产生了不利影响。第二种策略使得位点特异性切割成为可能，但是已经报道了过早切割，并且切割条件没有证明与活性生物分子的相容性。此外，可裂解连接体的一般适用性通常受到实施前需要多步有机合成的限制。腙已被探索用于生物分子的可逆缀合[5]和标记[6-8]。然而，它们的应用受到限制，因为大多数腙易于水解和过早裂解，而在生物和弱酸性条件下完全稳定的腙（和肟）水解较慢，并且难以交换或裂解。原则上，分子催化剂可用于加速水解并使
The selective enrichment of tagged molecules from complex biological mixtures is of primary importance for methods in chemical biology and proteomics. Affinity purification on (strept) avidin beads using biotin as an affinity tag is one of the most widely applied methods to achieve this, fully utilizing the high affinity of biotin for (strept) avidin (Ka≈ 1.7× 1015 M− 1)[1]. However, the method is limited by the harsh, denaturing conditions required for elution, such as an SDS boil or treatment with 8 M Guanidine (pH 1.5). Protein structure and function are lost and target-proteins may be contaminated with proteins nonspecifically bound to the beads. Two strategies have been explored to elute under mild conditions: i) weakening of the biotin-(strept) avidin interaction by modulating the Ka[2] and ii) introduction of a proteolytically [3] or chemically [4] cleavable linker. While the first strategy does improve the release of biotinylated proteins from (strept) avidin beads, it adversely affects the stringency of the immobilization. The second strategy enables site-specific cleavage, but premature cleavage has been reported and cleavage conditions have no demonstrated compatibility with active biomolecules. In addition, the general applicability of the cleavable linkers is often limited by the need for multistep organic synthesis before implementation.Hydrazones have been explored for the reversible conjugation [5] and labeling [6-8] of biomolecules. However, their application has been limited as most hydrazones are prone to hydrolysis and premature cleavage, while hydrazones (and oximes) that are fully stable under biological and mildly acidic conditions hydrolyze slower and are difficult to exchange or cleave. In principle a molecular catalyst could be used to accelerate hydrolysis and enable
登录
查看更多内容
影响因子：
15
作者：
Takaoka, Y;Tsutsumi, H;Hamachi, I
通讯作者：
Hamachi, I
影响因子：
15
作者：
Speers, AE;Cravatt, BF
通讯作者：
Cravatt, BF
影响因子：
48
作者：
Howarth, M;Chinnapen, DJF;Ting, AY
通讯作者：
Ting, AY
影响因子：
15
作者：
Dirksen, Anouk;Dirksen, Sjoerd;Dawson, Philip E.
通讯作者：
Dawson, Philip E.
影响因子：
37.8
作者：
Liu, D;Guo, H;Zlokovic, BV
通讯作者：
Zlokovic, BV