Engineering the Composition of Heterogeneous Lipid Bilayers to Stabilize Tethered Enzymes
Engineering the Composition of Heterogeneous Lipid Bilayers to Stabilize Tethered Enzymes
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DOI：
10.1002/admi.202000533
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发表时间：
2020-07-08
影响因子：
5.4
通讯作者：
Schwartz, Daniel K.
中科院分区：
文献类型：
--
作者：
Chaparro Sosa, Andres F.;Black, Kenneth J.;Schwartz, Daniel K.
脂双层作为酶的固定化表面，具有分子伴侣的活性，可以大大提高酶的稳定性。在这里，它表明，这种稳定效果可以推广到广泛的酶通过修改的混合脂质组合物的基础上的酶的性质。这通过分析硝基还原酶、脂肪酶、有机磷水解酶和溶菌酶在LB上的稳定性来证明，所述LB通过将两性离子1，2-二油酰基-sn-甘油基-3-磷酸胆碱与阴离子1，2-二油酰基-sn-甘油基-3-磷酸-（1 '-rac-甘油）或阳离子1，2-二油酰基-sn-甘油基-3-乙基磷酸胆碱混合而构成。有趣的是，虽然最佳脂质组合物表现出与每种酶具有相同符号的zeta电位，但稳定组成范围很宽，并且混合脂质LB比具有相似电荷的均匀LB更稳定。这表明，净电荷和成分的异质性都是重要的因素。此外，在极端变性条件下（7 murea）的适当的双层的稳定程度是前所未有的，和酶的活性是高达三倍以上的可溶性酶在每种酶的最佳温度。通过相关的酶扩散LB与来自单分子测量的重折叠动力学的信息，电荷和组合物对酶LB相互作用的影响，提供了一个基本原理的稳定作用的异质LB。
As surfaces for enzyme immobilization, lipid bilayers (LBs) have the potential to exhibit chaperone-like activity, which can greatly enhance enzyme stability. Here, it is shown that this stabilizing effect can be generalized to a broad range of enzymes by modifying the mixed lipid composition based on the properties of the enzyme. This is demonstrated by analyzing the stability of nitroreductase, lipase, organophosphorus hydrolase, and lysozyme on LBs composed by mixing zwitterionic 1,2-dioleoyl-sn-glycero-3-phosphocholine with anionic 1,2-dioleoyl-sn-glycero-3-phospho-(1 '-rac-glycerol) or cationic 1,2-dioleoyl-sn-glycero-3-ethylphosphocholine. Interestingly, while the optimal lipid composition exhibits a zeta potential with the same sign as each enzyme, the stabilizing compositional ranges are broad, and mixed lipid LBs are more stabilizing than homogeneous LBs with similar charge. This suggests that net charge and compositional heterogeneity are both important factors. Moreover, the degree of stabilization on appropriate bilayers under extremely denaturing conditions (7murea) is unprecedented, and activity of the enzymes is up to three-fold greater than that of the soluble enzyme at each enzyme's temperature optimum. By correlating enzyme diffusion on LBs with information about re-folding kinetics derived from single-molecule measurements, the influence of charge and composition on enzyme-LB interactions is shown, providing a rationale for the stabilizing effect of heterogeneous LBs.