Wang Deyi, Zhou Lingying, Zhang Xiaokun, Zhou Zixiang, Huang Zehuan, Gao Ning
School of Chemistry and Chemical Engineering, Beijing Institute of Technology, Beijing, 100081, P.R. China.
Key Laboratory of Polymer Chemistry and Physics of Ministry of Education, School of Materials Science and Engineering, Peking University, Beijing, 100871, P.R. China.
Angew Chem Int Ed Engl. 2025 Apr 1;64(14):e202422601. doi: 10.1002/anie.202422601. Epub 2025 Feb 3.
Dynamic liquid-liquid phase separation (LLPS) of intrinsically disordered proteins (IDPs) and associated assembly and disassembly of biomolecular condensates play crucial roles in cellular organization and metabolic networks. These processes are often regulated by supramolecular interactions. However, the complex and disordered structures of IDPs, coupled with their rapid conformational fluctuations, pose significant challenges for reconstructing supramolecularly-regulated dynamic LLPS systems and quantitatively illustrating variations in molecular interactions. Inspired by the structural feature of IDPs that facilitates LLPS, we designed a simplified phase-separating molecule, Nap-o-Nap, consisting of two naphthalene moieties linked by an ethylene glycol derivative. This compound exhibits LLPS under physiological conditions, forming coacervate microdroplets that undergo multiple cycles of disassembly and reassembly upon stoichiometric addition of Cucurbit[7]uril and Adamantane, respectively, based upon competitive host-guest interactions. Importantly, such reversible control offers a unique route to quantify entropically dominant nature (ΔS=14.0 cal ⋅ mol ⋅ K) within the LLPS process, in which the binding affinity of host-guest interactions (ΔG=-14.9 kcal ⋅ mol) surpass that of the LLPS of Nap-o-Nap (ΔG=-2.1 kcal ⋅ mol), enabling the supramolecular regulation process. The supramolecularly switched LLPS, along with selective client recruitment and exclusion by resultant coacervates, provides a promising platform for either boosting or retarding enzymatic reactions, thereby orchestrating biological enzyme kinetics.
内在无序蛋白（IDP）的动态液-液相分离（LLPS）以及生物分子凝聚物的相关组装和解聚在细胞组织和代谢网络中起着关键作用。这些过程通常由超分子相互作用调控。然而，IDP复杂且无序的结构，加上其快速的构象波动，为重建超分子调控的动态LLPS系统以及定量说明分子相互作用的变化带来了重大挑战。受IDP促进LLPS的结构特征启发，我们设计了一种简化的相分离分子Nap-o-Nap，它由两个通过乙二醇衍生物连接的萘部分组成。该化合物在生理条件下表现出LLPS，形成凝聚微滴，基于竞争性主客体相互作用，分别在化学计量添加葫芦[7]脲和金刚烷时经历多次解聚和重新组装循环。重要的是，这种可逆控制为量化LLPS过程中熵主导性质（ΔS = 14.0 cal·mol·K）提供了一条独特途径，其中主客体相互作用的结合亲和力（ΔG = -14.9 kcal·mol）超过了Nap-o-Nap的LLPS的结合亲和力（ΔG = -2.1 kcal·mol），从而实现超分子调控过程。超分子开关的LLPS，以及由此产生的凝聚物对选择性客体的招募和排斥，为加速或延缓酶促反应提供了一个有前景的平台，从而调控生物酶动力学。
