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. 2022 Jan 24;18(9):2020–2035. doi: 10.1080/15548627.2022.2025572

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Figure 5. — Protein chemical synthesis strategies to produce LC3B–PE in vitro. (A) PolyArg-assisted solubilization strategy to produce LC3B–PE. Recombinant LC3B[1-114] MESNa thioester is obtained by an intein strategy. LC3B[115-120] peptide CQETFG containing DSPE fused with a polyArg tag is obtained by SPPS. LC3B[1-114] MESNa thioester and Arg4-tagged peptide-DSPE are conjugated by EPL. Following photosensitive Arg4-tag removal, LC3B–DSPE proteins are produced. (B) MBP-assisted solubilization strategy to produce LC3B–PE. Recombinant LC3B[1-114] MESNa thioester fused with an MBP tag is obtained by an intein strategy. LC3B[115-120] peptide CQETFG containing DPPE is produced by SPPS. LC3B[1-114] MESNa thioester and peptide-DPPE are conjugated by EPL. Following TEV protease cleavage, LC3B–DPPE proteins are produced.