Intracellular Biomacromolecule Delivery by Stimuli-Responsive Protein Vesicles Loaded by Hydrophobic Ion Pairing

ACS Omega, Год журнала: 2025, Номер unknown

Опубликована: Янв. 14, 2025

Proteins can perform ideal therapeutic functions. However, their large size and significant surface hydrophilicity charge prohibit them from reaching intracellular targets. These chemical features also render poorly encapsulated by nanoparticles used for delivery. In this work, a novel combination of protein vesicles hydrophobic ion pairing (HIP) was to load cargo achieve cytosolic delivery overcome the limitations previous vesicle properties. Protein are thermally self-assembling made elastin-like polypeptide (ELP) fused an arginine-rich leucine zipper globular glutamate-rich zipper. To impart stimuli-responsive disassembly, physiological stability, small size, ELP sequence modified include histidine tyrosine residues. HIP release requiring endosomal escape function. enabled cytochrome c, cytosolically active protein, reduction in viability both traditional two-dimensional (2D) human cancer cell line culture biomimetic three-dimensional (3D) organoid model acute myeloid leukemia. By examining uptake positively negatively charged fluorescent cargos loaded HIP, work revealed necessity how loading influences self-assembly disassembly using microscopy, small-angle X-ray scattering, nanoparticle tracking analysis. have potential broaden use proteins as therapeutics various diseases extend deliver other biomacromolecules, strategy developed here resulted first vesicles.

Язык: Английский

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Design of a diblock-based membraneless organelle system for metabolic process control

Chemical Engineering Journal, Год журнала: 2025, Номер unknown, С. 160239 - 160239

Опубликована: Фев. 1, 2025

Язык: Английский

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Sequence-defined structural transitions by calcium-responsive proteins

Опубликована: Авг. 20, 2024

Biopolymer sequences dictate their functions, and protein-based polymers are a promising platform to establish sequence–function relationships for novel biopolymers. To efficiently explore vast sequence spaces of natural proteins, repetition is common strategy tune amplify specific functions. This applied repeats-in-toxin (RTX) proteins with calcium-responsive folding behavior, which stems from tandem repeats the nonapeptide GGXGXDXUX in X can be any amino acid U hydrophobic acid. determine functional range this nonapeptide, we modified naturally occurring RTX protein that forms β-roll structures presence calcium. Sequence modifications focused on calcium-binding turns within repetitive region, including either global substitution nonconserved residues or complete replacement consensus GGAGXDTLY. Some disrupted typical transition intrinsically disordered random coils folded β rolls, despite conservation underlying sequence. Proteins enriched smaller, acids adopted secondary absence calcium underwent structural rearrangement calcium-rich environments. In contrast, bulkier, hydrophilic maintained intrinsic disorder These results indicate significant role folding, thereby revealing leverage design tunable,

Язык: Английский

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Sequence-defined structural transitions by calcium-responsive proteins

Polymer Chemistry, Год журнала: 2024, Номер 15(47), С. 4864 - 4874

Опубликована: Янв. 1, 2024

Recombinant protein engineering accelerated the synthesis of sequence-defined, calcium-responsive biopolymers. A mutation panel repeats-in-toxin (RTX) proteins revealed sequence-dependent disorder, calcium sensitivity, and structural transitions.

Язык: Английский

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