Department of Chemistry, University of British Columbia, Vancouver, British Columbia, Canada.
Biophys J. 2011 Apr 6;100(7):1794-9. doi: 10.1016/j.bpj.2011.02.030.
Rationally enhancing the mechanical stability of proteins remains a challenge in the field of single molecule force spectroscopy. Here we demonstrate that it is feasible to use a "cocktail" approach for combining more than one approach to enhance significantly the mechanical stability of proteins in an additive fashion. As a proof of principle, we show that metal chelation and protein-protein interaction can be combined to enhance the unfolding force of a protein to ∼450 pN, which is >3 times of its original value. This is also higher than the mechanical stability of most of proteins studied so far. We also extend such a cocktail concept to combine two different metal chelation sites to enhance protein mechanical stability. This approach opens new avenues to efficiently regulating the mechanical properties of proteins, and should be applicable to a wide range of elastomeric proteins.
理性地增强蛋白质的机械稳定性仍然是单分子力谱学领域的一个挑战。在这里，我们证明了使用“鸡尾酒”方法将一种以上的方法结合起来以附加的方式显著提高蛋白质的机械稳定性是可行的。作为原理的证明，我们表明可以将金属螯合和蛋白质-蛋白质相互作用结合起来，将蛋白质的展开力提高到约 450 pN，这是其原始值的 3 倍以上。这也高于迄今为止研究的大多数蛋白质的机械稳定性。我们还将这种鸡尾酒概念扩展到结合两个不同的金属螯合位点来增强蛋白质的机械稳定性。这种方法为有效地调节蛋白质的机械性能开辟了新的途径，并且应该适用于广泛的弹性蛋白。
