Chemical Engineering Journal, Journal Year: 2025, Volume and Issue: unknown, P. 160239 - 160239
Published: Feb. 1, 2025
Language: Английский
Nature Chemical Biology, Journal Year: 2024, Volume and Issue: 20(11), P. 1420 - 1433
Published: Sept. 26, 2024
Language: Английский
Citations
10
bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown
Published: Jan. 2, 2025
Abstract Proteins commonly self-assemble to create liquid or solid condensates with diverse biological activities. The mechanisms of assembly are determined by each protein’s sequence and cellular context. We previously developed distributed amphifluoric FRET (DAmFRET) analyze determinants self-assembly in cells. Here, we extend DAmFRET creating a nanobody (mEosNb) against the fluorescent protein mEos3 physically tether thereby recruit candidate modifier proteins mEos3-fused query proteins. This accessorization allows us rapidly screen for effects on phase behavior modulating expression level valency mEosNb-fused modifiers. show that our system recapitulates known multivalency liquid-liquid separation can discriminate between nucleation amyloid amyloid-like assemblies. Our approach adds new experimental dimension interrogating intracellular transitions. Lay summary Protein self-assemblies essential function, but also contribute disease. develop tool study how their formation is influenced other factors. control location number interactions interest may influence it. results provide insight into will aid research toward treating diseases associated aberrant assembly.
Language: Английский
Citations
1
Nature reviews. Immunology, Journal Year: 2025, Volume and Issue: unknown
Published: Jan. 28, 2025
Language: Английский
Citations
1
Nature Chemistry, Journal Year: 2025, Volume and Issue: unknown
Published: March 12, 2025
Language: Английский
Citations
1
Trends in Biochemical Sciences, Journal Year: 2024, Volume and Issue: 49(7), P. 633 - 648
Published: April 22, 2024
Language: Английский
Citations
7
ChemCatChem, Journal Year: 2024, Volume and Issue: unknown
Published: May 8, 2024
Abstract Biomolecular condensates are membraneless organelles that orchestrate various metabolic pathways in living cells. Understanding how these crowded structures regulate enzyme reactions remains yet challenging due to their dynamic and intricate nature. Coacervate microdroplets formed by associative liquid‐liquid phase separation of oppositely charged polyions have emerged as relevant condensate models study catalysis. Enzyme within droplets show altered kinetics, influenced factors such substrate partitioning, crowding, interactions with coacervate components; it is often disentangle the contributions each. Here, we investigate peroxidase activity a de novo polysaccharide‐based coacervates. By comparing reaction kinetics buffer, suspension coacervates bulk collected after centrifugation droplets, significantly increases catalytic efficiency. We demonstrate main origin this enhanced lies macromolecular crowding coupled changes conformational dynamics environment. Altogether, findings underline crucial role matrix catalysis, beyond simple partitioning effects. The observed boost provides insights for designing biocatalytically active synthetic organelles.
Language: Английский
Citations
6
Small, Journal Year: 2024, Volume and Issue: unknown
Published: May 28, 2024
Abstract Membraneless organelles are cellular biomolecular condensates that formed by liquid–liquid phase separation (LLPS) of proteins and nucleic acids. LLPS is driven multiple weak attractive forces, including intermolecular interactions mediated aromatic amino Considering the contribution π‐electron bearing side chains to protein‐RNA LLPS, systematically study sought how composition acids affects formation heterotypic their physical properties. For this, a library minimalistic peptide building blocks designed containing varying number compositions It shown aromatics in sequence affect propensity, material properties (bio)chemical stability peptide/RNA condensates. The findings shed light on aromatics’ These insights can be applied for regulation condensate improvement stability, various biomedical biotechnological applications.
Language: Английский
Citations
6
Journal of Chemical Theory and Computation, Journal Year: 2024, Volume and Issue: 20(15), P. 6881 - 6889
Published: July 30, 2024
Biomolecules composed of a limited set chemical building blocks can colocalize into distinct, spatially segregated compartments known as biomolecular condensates. While many condensates are to form spontaneously via phase separation, it has been unclear how immiscible with precisely controlled molecular compositions assemble from small number blocks. We address this question by establishing connection between the specificity interactions and thermodynamic stability coexisting By computing minimum interaction required target compositions, we show design heteropolymer mixtures that produce compositionally complex using only monomer types. Our results provide insight compositional arises in naturally occurring multicomponent demonstrate rational algorithm for engineering artificial simple
Language: Английский
Citations
6
Biomacromolecules, Journal Year: 2024, Volume and Issue: 25(4), P. 2449 - 2461
Published: March 14, 2024
Peptide-based materials are diverse candidates for self-assembly into modularly designed and stimuli-responsive nanostructures with precisely tunable compositions. Here, we genetically fused computationally coiled coil-forming peptides to the N- C-termini of compositionally distinct multistimuli-responsive resilin-like polypeptides (RLPs) various lengths. The successful expression these hybrid in bacterial hosts was confirmed through techniques such as gel electrophoresis, mass spectrometry, amino acid analysis. Circular dichroism spectroscopy ultraviolet–visible turbidimetry demonstrated that despite fusion disparate structural responsive units, coils remained stable polypeptides, sequence-encoded differences thermoresponsive phase separation RLPs were preserved. Cryogenic transmission electron microscopy coarse-grained modeling showed after thermal annealing solution, adopted a closed loop conformation assembled nanofibrils capable further hierarchically organizing cluster structures ribbon-like mediated by self-association tendency RLPs.
Language: Английский
Citations
5
ACS Nano, Journal Year: 2024, Volume and Issue: 18(26), P. 16530 - 16544
Published: June 14, 2024
The cellular environment, characterized by its intricate composition and spatial organization, hosts a variety of organelles, ranging from membrane-bound ones to membraneless structures that are formed through liquid–liquid phase separation. Cells show precise control over the position such condensates. We demonstrate organelle movement in external concentration gradients, diffusiophoresis, is distinct one colloids because fluxes can remain finite inside liquid-phase droplets latter arises incompressibility. Within domains diffusiophoresis naturally biochemical reactions driven chemical fuel produce waste. Simulations analytical arguments within minimal model reaction-driven separation reveal directed stems two contributions: Fuel waste refilled or extracted at boundary, resulting which (i) induce product via incompressibility (ii) result an asymmetric forward reaction droplet's surroundings (as well as backward droplet), thereby shifting position. former contribution dominates sets direction movement, toward away source sink, depending on molecules' affinity waste, respectively. mechanism thus provides simple means organize condensates with different composition. Particle-based simulations systems more complex cycles corroborate robustness universality this mechanism.
Language: Английский
Citations
5