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Intracellular artificial supramolecules based on de novo designed Y15 peptides.

ABSTRACT: De novo designed self-assembling peptides (SAPs) are promising building blocks of supramolecular biomaterials, which can fulfill a wide range of applications, such as scaffolds for tissue culture, three-dimensional cell culture, and vaccine adjuvants. Nevertheless, the use of SAPs in intracellular spaces has mostly been unexplored. Here, we report a self-assembling peptide, Y15 (YEYKYEYKYEYKYEY), which readily forms β-sheet structures to facilitate bottom-up synthesis of functional protein assemblies in living cells. Superfolder green fluorescent protein (sfGFP) fused to Y15 assembles into fibrils and is observed as fluorescent puncta in mammalian cells. Y15 self-assembly is validated by fluorescence anisotropy and pull-down assays. By using the Y15 platform, we demonstrate intracellular reconstitution of Nck assembly, a Src-homology 2 and 3 domain-containing adaptor protein. The artificial clusters of Nck induce N-WASP (neural Wiskott-Aldrich syndrome protein)-mediated actin polymerization, and the functional importance of Nck domain valency and density is evaluated.

SUBMITTER: Miki T

PROVIDER: S-EPMC8185068 | biostudies-literature |

REPOSITORIES: biostudies-literature

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De novo designed transmembrane peptides activating the α5β1 integrin.

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| S-EPMC6151875 | biostudies-literature

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Metal Stabilization of Collagen and de Novo Designed Mimetic Peptides.

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| S-EPMC5335877 | biostudies-literature

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| S-EPMC5019575 | biostudies-literature

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| S-EPMC3502156 | biostudies-literature

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| S-EPMC9495127 | biostudies-literature

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| S-EPMC9979648 | biostudies-literature

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