Design of Multimodal Supramolecular Protein Assemblies via Enzyme–Substrate Interactions for Intracellular Antioxidant Regulation

Nano Letters, Journal Year: 2025, Volume and Issue: unknown

Published: March 11, 2025

Allosteric modulation of protein function, which involves effector binding triggering distant conformational changes, is crucial for cellular and metabolic control. However, achieving tunable control, structural diversity, precise intracellular regulation remains challenging. Here, we designed dynamic supramolecular assemblies driven by enzyme-substrate interactions antioxidant in cells. Using a glutathione S-transferase modified with cysteine mutation (GSTK77C), engineered an molecule (GMP4M) containing (GSH) moiety maleimide group linked PEG chain. This system forms hierarchical diverse morphologies, including nanowires, nanorings, nanobranches, nanotwists, switchable "ON/OFF" enzymatic activity modulated endogenous GSH. The maintain integrity under physiological conditions, show remarkable reversibility, outperform native GST stability environmental adaptability. approach provides versatile platform creating broad applications therapies biomedical interventions.

Language: Английский

Molecular Design of Encapsulin Protein Nanoparticles to Display Rotavirus Antigens for Enhancing Immunogenicity

Vaccines, Journal Year: 2024, Volume and Issue: 12(9), P. 1020 - 1020

Published: Sept. 6, 2024

Rotavirus considerably threatens global health, particularly for children <5 years. Current, licensed oral attenuated vaccine formulations have limitations including insufficient efficacy in low- and middle-income countries, warranting urgent development of novel vaccines with improved safety profiles. Herein, we present a approach utilizing an encapsulin (ENC) nanoparticle (NP)-based non-replicating rotavirus vaccine. ENC, originating from bacteria, offers self-assembling scaffold that displays VP8* antigens on its surface. To enhance the correct folding soluble expression monomeric their subsequent assembly into NP, adopted RNA-interacting domain (RID) mammalian transfer RNA synthetase as tag fused to N-terminus ENC-VP8* fusion protein. Using RID-ENC-VP8* tripartite modular design, insertion linkers appropriate length sequence universal T cell epitope P2 remarkably production yield immunogenicity. Cleavage RID rendered homogenous ENC-P2-VP8* protein NPs. Immunization induced markedly higher levels VP8*-specific antibodies virus neutralization titers mice than those by P2-VP8* without ENC. Altogether, these results highlight potential designed ENC NP-based effective strategy against disease address health challenges.

Language: Английский

Genetically Engineered Liposwitch-Based Nanomaterials

Biomacromolecules, Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 4, 2024

Fusion of intrinsically disordered and globular proteins is a powerful strategy to create functional nanomaterials. However, the immutable nature genetic encoding restricts dynamic adaptability nanostructures postexpression. To address this, we envisioned using myristoyl switch, protein that combines allostery post-translational modifications─two strategies modify properties without altering their sequence─to regulate (IDP)-driven nanoassembly. A typical allosterically activated by stimulus, reveals sequestered lipid for membrane association. We hypothesize this conditional exposure lipids can assembly fusion proteins, concept term "liposwitching". tested fusing recoverin, calcium-dependent with elastin-like polypeptide, thermoresponsive model IDP. Biophysical analyses confirmed recoverin's myristoyl-switch functionality, while light scattering cryo-transmission electron microscopy showed distinct calcium- lipidation-dependent phase separation assembly. This study highlights liposwitching as viable controlling DP-driven nanoassembly, enabling applications in synthetic biology cellular engineering.

Language: Английский