Self-Assembling Polypeptides in Complex Coacervation

Autor(es)

Sathyavageeswaran, Arvind ; Sabadini, Julia Bonesso ; Perry, Sarah L.

• Tipo de documento: Artigo Científico
• Fonte: ACCOUNTS OF CHEMICAL RESEARCH; v. 57, n. 3, p. 13-pg., 2024-01-22.

Resumo

Intracellular compartmentalization plays a pivotal role in cellular function, with membrane-bound organelles and membrane-less biomolecular "condensates" playing key roles. These condensates, formed through liquid-liquid phase separation (LLPS), enable selective compartmentalization without the barrier of a lipid bilayer, thereby facilitating rapid formation and dissolution in response to stimuli. Intrinsically disordered proteins (IDPs) or proteins with intrinsically disordered regions (IDRs), which are often rich in charged and polar amino acid sequences, scaffold many condensates, often in conjunction with RNA. Comprehending the impact of IDP/IDR sequences on phase separation poses a challenge due to the extensive chemical diversity resulting from the myriad amino acids and post-translational modifications. To tackle this hurdle, one approach has been to investigate LLPS in simplified polypeptide systems, which offer a narrower scope within the chemical space for exploration. This strategy is supported by studies that have demonstrated how IDP function can largely be understood based on general chemical features, such as clusters or patterns of charged amino acids, rather than residue-level effects, and the ways in which these kinds of motifs give rise to an ensemble of conformations. Our laboratory has utilized complex coacervates assembled from oppositely charged polypeptides as a simplified material analogue to the complexity of liquid-liquid phase separated biological condensates. Complex coacervation is an associative LLPS that occurs due to the electrostatic complexation of oppositely charged macro-ions. This process is believed to be driven by the entropic gains resulting from the release of bound counterions and the reorganization of water upon complex formation. Apart from their direct applicability to IDPs, polypeptides also serve as excellent model polymers for investigating molecular interactions due to the wide range of available side-chain functionalities and the capacity to finely regulate their sequence, thus enabling precise control over interactions with guest molecules. Here, we discuss fundamental studies examining how charge patterning, hydrophobicity, chirality, and architecture affect the phase separation of polypeptide-based complex coacervates. These efforts have leveraged a combination of experimental and computational approaches that provide insight into molecular level interactions. We also examine how these parameters affect the ability of complex coacervates to incorporate globular proteins and viruses. These efforts couple directly with our fundamental studies into coacervate formation, as such "guest" molecules should not be considered as experiencing simple encapsulation and are instead active participants in the electrostatic assembly of coacervate materials. Interestingly, we observed trends in the incorporation of proteins and viruses into coacervates formed using different chain length polypeptides that are not well explained by simple electrostatic arguments and may be the result of more complex interactions between globular and polymeric species. Additionally, we describe experimental evidence supporting the potential for complex coacervates to improve the thermal stability of embedded biomolecules, such as viral vaccines. Ultimately, peptide-based coacervates have the potential to help unravel the physics behind biological condensates, while paving the way for innovative methods in compartmentalization, purification, and biomolecule stabilization. These advancements could have implications spanning medicine to biocatalysis. (AU)

• Processo FAPESP: 18/25041-5 - Avaliação do estado de equilíbrio e estrutura de coacervados formados por copolímeros-em-bloco e polímeros de carga oposta
• Beneficiário: Julia Bonesso Sabadini
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 21/12071-6 - Arquitetando coloides via interações supramoleculares: de fundamentos a aplicações
• Beneficiário: Watson Loh
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 15/25406-5 - Organizando a matéria: colóides formados por associação de surfactantes, polímeros e nanopartículas
• Beneficiário: Watson Loh
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 20/11735-5 - Micelas de polieletrólitos: caracterização, propriedades e aplicações para a incorporação de enzimas
• Beneficiário: Julia Bonesso Sabadini
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto
• Processo FAPESP: 21/11317-1 - Design de regras para o encapsulamento de proteínas no interior de micelas de núcleo coacervado (C3M)
• Beneficiário: Julia Bonesso Sabadini
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Doutorado Direto