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TitleCell-Free Nonequilibrium Assembly for Hierarchical Protein/Peptide Nanopillars.
Journal, issue, pagesJ Am Chem Soc, Vol. 146, Issue 38, Page 26102-26112, Year 2024
Publish dateSep 25, 2024
AuthorsJiaqi Guo / Ayisha Zia / Qianfeng Qiu / Michael Norton / Kangqiang Qiu / Junichi Usuba / Zhiyu Liu / Meihui Yi / Shane T Rich-New / Michael Hagan / Seth Fraden / Grace D Han / Jiajie Diao / Fengbin Wang / Bing Xu /
PubMed AbstractCells contain intricate protein nanostructures, but replicating them outside of cells presents challenges. One such example is the vertical fibronectin pillars observed in embryos. Here, we ...Cells contain intricate protein nanostructures, but replicating them outside of cells presents challenges. One such example is the vertical fibronectin pillars observed in embryos. Here, we demonstrate the creation of cell-free vertical fibronectin pillar mimics using nonequilibrium self-assembly. Our approach utilizes enzyme-responsive phosphopeptides that assemble into nanotubes. Enzyme action triggers shape changes in peptide assemblies, driving the vertical growth of protein nanopillars into bundles. These bundles, with peptide nanotubes serving as a template to remodel fibronectin, can then recruit collagen, which forms aggregates or bundles depending on their types. Nanopillar formation relies on enzyme-catalyzed nonequilibrium self-assembly and is governed by the concentrations of enzyme, protein, peptide, the structure of the peptide, and peptide assembly morphologies. Cryo-EM reveals unexpected nanotube thinning and packing after dephosphorylation, indicating a complex sculpting process during assembly. Our study demonstrates a cell-free method for constructing intricate, multiprotein nanostructures with directionality and composition.
External linksJ Am Chem Soc / PubMed:39255453
MethodsEM (helical sym.)
Resolution2.8 - 3.0 Å
Structure data

44636

9bjn

EMDB-44636, PDB-9bjn:
Cryo-EM of Azo-ffspy fiber
Method: EM (helical sym.) / Resolution: 3.0 Å

44637

9bjo

EMDB-44637, PDB-9bjo:
Cryo-EM of Azo-ffsy fiber
Method: EM (helical sym.) / Resolution: 2.8 Å

Source
  • synthetic construct (others)
KeywordsPROTEIN FIBRIL / D-peptide / peptide-fiber / helical

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