Spatiotemporal Dynamic Assembly/Disassembly of Organelle‐Mimics Based on Intrinsically Disordered Protein‐Polymer Conjugates

Abstract

Design of reversible organelle‐like microcompartments formed by liquid–liquid phase separation in cell‐mimicking entities has significantly advanced the bottom‐up construction of artificial eukaryotic cells. However, organizing the formation of artificial organelle architectures in a spatiotemporal manner within complex primitive compartments remains scarcely explored. In this work, thermoresponsive hybrid polypeptide‐polymer conjugates are rationally engineered and synthesized, resulting from the conjugation of an intrinsically disordered synthetic protein (IDP), namely elastin‐like polypeptide, and synthetic polymers (poly(ethylene glycol) and dextran) that are widely used as macromolecular crowding agents. Cell‐like constructs are built using droplet‐based microfluidics that are filled with such bioconjugates and an artificial cytoplasm system that is composed of specific polymers conjugated to the IDP. The distinct spatial organizations of two polypeptide‐polymer conjugates and the dynamic assembly and disassembly of polypeptide‐polymer coacervate droplets in response to temperature are studied in the cytomimetic protocells. Furthermore, a monoblock IDP with longer length is concurrently included with bioconjugates individually inside cytomimetic compartments. Both bioconjugates exhibit an identical surfactant‐like property, compartmentalizing the monoblock IDP coacervates via temperature control. These findings lay the foundation for developing hierarchically structured synthetic cells with interior organelle‐like structures which could be designed to localize in desired phase‐separated subcompartments.

Ultraslow Biological Water-Like Dynamics in Waterless Liquid Protein

“…4 Their potential is immense for applications like drug delivery, 5 biosensing, and biomimetics, 6,7 as universal solvents, 8 and in the design of organelle mimics. 9 Globular proteins retain their structure and function, while intrinsically disordered proteins (IDPs) can adopt a secondary structure in a waterless environment. 10 Such unusual phenomena, arising as a result of the crowded environment in waterless protein liquids, make them good models for crowded biological systems, in which proteins often behave differently from their dilute solutions.…”

“…The viscous solvent-free/waterless protein liquids, obtained upon removal of water from these assemblies, exhibit unusually high thermal stability, unique ability to extend the range of operation of enzymes to temperatures as high as 150 °C, and enhanced enzyme activity . Their potential is immense for applications like drug delivery, biosensing, and biomimetics, , as universal solvents, and in the design of organelle mimics . Globular proteins retain their structure and function, while intrinsically disordered proteins (IDPs) can adopt a secondary structure in a waterless environment .…”

Peptide–Polymer Conjugates: A Promising Therapeutic Solution for Drug-Resistant Bacteria

“…For example, when AMPs is combined with biocompatible polymers, its positive charge can be partially neutralized, so as to help AMPs escape the attack of the immune system, improve its in vivo stability, and prolong its in vivo circulation time. Meanwhile, the cytotoxicity of AMPs was also reduced [65][66][67].…”

Sequence Control of the Self-Assembly of Elastin-Like Polypeptides into Hydrogels with Bespoke Viscoelastic and Structural Properties

“…This has inspired the development of elastin-like polypeptides (ELPs) and derivatives thereof for the manufacture of dynamic and thermoresponsive materials in fields like tissue engineering, drug delivery, microfluidics, and actuation . ELPs have been used also for the development of micelles, vesicles, artificial cells, membraneless organelles, and as models of intrinsically disordered proteins. ,− Like tropoelastin, ELP solutions also display LCST phase behavior. , ELPs are intrinsically disordered and highly dynamic both below and above the T t . , Above the T t , ELP–ELP interactions become favored over ELP–solvent interactions, and thus the solution segregates into ELP-rich and solvent-rich phases . The thermoresponsiveness of ELPs can be controlled by sequence design (e.g., block arrangement or molecular weight) or by processing conditions (e.g., protein concentration or solution conditions), and numerous studies have shown the ability of ELPs to form a rich landscape of sophisticated nanostructures above the T t . − …”