A Balance Between Capture and Release: How Nanochaperones Regulate Refolding of Thermally Denatured Proteins

Abstract

Nanochaperones have been designed and used for regulating the (re)folding of proteins, treating protein misfolding‐related diseases, and, more recently, in drug delivery. Despite various successes, a complete understanding of the working mechanisms remains elusive, which represents a challenge for the realization of their full potential. Here, we thoroughly investigated the functioning of differently charged nanochaperones that regulate the refolding of thermally denatured lysozyme. We found that the balance between the capture and release of lysozyme clients that are controlled by nanochaperones plays a key role in regulating refolding. More importantly, the findings could be applied to other enzymes with various physicochemical properties. On the basis of these results, we could recover the activity of enzymes with high efficiency either after 20 days of storage at 40 °C or heating at high temperatures for 30–60 min. Our results provide important new design strategies for nanochaperone systems to improve their properties and expand their applications.

Tailoring a Nanochaperone to Regulate α‐Synuclein Assembly

“…This is a prerequisite for the prophylactic use in α‐synucleinopathy because tight capture of α‐Syn monomer would affect its normal function. In our previous work, the balance between the capture and release of different protein targets could be regulated by adjusting the ratio of hydrophobic and electrostatic interactions [5c] . The solvent‐exposed hydrophobic surface of monomeric α‐Syn increases when it is transformed into toxic oligomers, [7a] which is similar to the properties of folded and unfolded protein.…”

One-pot synthesis of high-concentration mixed-shell polymeric micelles as nanochaperones for the renaturation of bulk proteins

“…Furthermore, the synthesized nanochaperones have considerable refolding capabilities compared to the previously reported nanochaperones by our group. 27,45 The synthetic nanochaperones developed here can be used to assist the refolding of bulk denatured proteins to their bioactive form, and stabilize native proteins for long-term storage.…”

“…[21][22][23] Inspired by natural chaperones, we have developed a series of self-assembled mixedshell polymeric micelle (MSPM) based nanochaperones and shown that they are efficient in regulating the activities of proteins, such as (re)folding, misfolding and aggregation. [24][25][26][27] The MSPMs are generally fabricated by the self-assembly of two or more amphiphilic di-block polymers in selective solvents. [28][29][30][31][32] However, this manufacturing method is complicated and time-consuming, and the concentration of the final products is low, which greatly restricts its large-scale applicability in practice.…”

Thermo-Gelling Dendronized Chitosans for Modulating Protein Activity

“…Biomacromolecules, including proteins and nucleic acids, often play pivotal roles during cellular biotransformation. , Generally, the regulation process of their activities is accomplished through the assistance of molecular chaperones in crowded microenvironments. , However, the lesion of their biological activity is unavoidable due to extracellular environments during their applications in diagnostics and therapeutics. , To improve the stability of proteins effectively, various synthetic or natural materials have been used as their artificial chaperones and combined with them to form the corresponding hybrid conjugates. , Typically, these combination processes are accomplished by embedding the active site of biomacromolecules into synthetic scaffolds through covalent and/or noncovalent interactions. , In particular, combination ways between proteins and scaffolds via supramolecular interactions, such as electrostatic interactions, are more flexible . This provides these conjugates with more possible adjustability (such as release capabilities) of biomacromolecules without destroying their bioactivity. , Notably, utilization of stimulus response macromolecules as the scaffold for proteins brings convenience for the regulation of their bioactivities and release processes of the proteins under designated conditions (for instance, temperature, pH, or degrading enzyme). , …”

“…2,6 Typically, these combination processes are accomplished by embedding the active site of biomacromolecules into synthetic scaffolds through covalent and/or noncovalent interactions. 5,7 In particular, combination ways between proteins and scaffolds via supramolecular interactions, such as electrostatic interactions, are more flexible. 8 This provides these conjugates with more possible adjustability (such as release capabilities) of biomacromolecules without destroying their bioactivity.…”