Key Laboratory of Functional Polymer Materials of Ministry of Education, State Key Laboratory of Medicinal Chemical Biology and Institute of Polymer Chemistry, College of Chemistry, Nankai University , Tianjin 300071, China.
ACS Nano. 2017 Oct 24;11(10):10549-10557. doi: 10.1021/acsnano.7b05947. Epub 2017 Oct 9.
The folding process of a protein is inherently error-prone, owing to the large number of possible conformations that a protein chain can adopt. Partially folded or misfolded proteins typically expose hydrophobic surfaces and tend to form dysfunctional protein aggregates. Therefore, materials that can stabilize unfolded proteins and then efficiently assist them refolding to its bioactive form are of significant interest. Inspired by natural chaperonins, we have synthesized a series of polymeric nanochaperones that can facilitate the refolding of denatured proteins with a high recovery efficiency (up to 97%). Such nanochaperones possess phase-separated structure with hydrophobic microdomains on the surface. This structure allows nanochaperones to stabilize denatured proteins by binding them to the hydrophobic microdomains. We have also investigated the mechanism by which nanochaperones assist the protein refolding and established the design principles of nanochaperones in order to achieve effective recovery of a certain protein from their denatured forms. With a carefully designed composition of the microdomains according to the surface properties of the client proteins, the binding affinity between the hydrophobic microdomain and the denatured protein molecules can be tuned precisely, which enables the self-sorting of the polypeptides and the refolding of the proteins into their bioactive states. This work provides a feasible and effective strategy to recover inclusion bodies to their bioactive forms, which has potential to reduce the cost of the manufacture of recombinant proteins significantly.
蛋白质的折叠过程本身容易出错，这是由于蛋白质链可以采用的构象数量众多。部分折叠或错误折叠的蛋白质通常会暴露出疏水面，并倾向于形成无功能的蛋白质聚集体。因此，能够稳定未折叠蛋白质并有效地帮助它们重折叠为其生物活性形式的材料具有重要意义。受天然伴侣蛋白的启发，我们合成了一系列聚合物纳米伴侣，它们可以以高回收率（高达 97%）促进变性蛋白质的重折叠。这些纳米伴侣具有相分离结构，表面上有疏水性微区。这种结构允许纳米伴侣通过与疏水性微区结合来稳定变性蛋白质。我们还研究了纳米伴侣辅助蛋白质重折叠的机制，并建立了纳米伴侣的设计原则，以实现从变性形式有效回收特定蛋白质。根据客户蛋白质的表面特性，精心设计微区的组成，可以精确调整疏水性微区与变性蛋白质分子之间的结合亲和力，从而实现多肽的自分类和蛋白质重折叠为其生物活性状态。这项工作为恢复包涵体的生物活性形式提供了一种可行且有效的策略，有潜力显著降低重组蛋白制造的成本。
