Recombinant Synthesis of Hybrid Lipid–Peptide Polymer Fusions that Self‐Assemble and Encapsulate Hydrophobic Drugs

Abstract

Inspired by biohybrid molecules that are synthesized in Nature through post-translational modification (PTM), we have exploited a eukaryotic PTM to recombinantly synthesize lipid-polypeptide hybrid materials. By co-expressing yeast N-myristoyltransferase with an elastin-like polypeptide (ELP) fused to a short recognition sequence in E. coli, we show robust and high yield modification of the ELP with myristic acid. The ELP’s reversible phase behavior is retained upon myristoylation and can be tuned to span a 30–60 °C. Myristoylated ELPs provide a versatile platform for genetically pre-programming self-assembly into micelles of varied size and shape. Their lipid cores can be loaded with hydrophobic small molecules by passive diffusion. Encapsulated doxorubicin and paclitaxel exhibit cytotoxic effects on 4T1 and PC3-luc cells, respectively, with potencies similar to chemically conjugated counterparts, and longer plasma circulation than free drug upon intravenous injection in mice.

Extracellular Elastin Molecule Modulates Alzheimer’s Aβ Dynamics In Vitro and In Vivo by Affecting Microglial Activities

“…34,35 The primary structure of ELPs consists of repetitive pentapeptide units (VPGVG) n with low complexity. [36][37][38] ELP90, which has 96 (VPGVG) repeats and a molecular weight of 39.68 kDa, was used in this study (Supporting Information Figure S1). The protein was purified by chromatography and characterized via polyacrylamide gel electrophoresis (PAGE) (Figure 1).…”

Significant Upregulation of Alzheimer's β‐Amyloid Levels in a Living System Induced by Extracellular Elastin Polypeptides

“…The ELPs are a family of genetically encoded biopolymers and contain repeats of canonical sequences derived from native elastin. The primary structure of ELPs consists of repetitive pentapeptide units (VPGVG)n with low complexity (Figure A) . ELP30, ELP48, and ELP90 with varied lengths of polypeptide backbones were used in this study.…”

Engineering the Architecture of Elastin‐Like Polypeptides: From Unimers to Hierarchical Self‐Assembly

“…Copyright 2012, American Chemical Society. (b) Reproduced with permission . Copyright 2017, John Wiley and Sons.…”

“…This approach was first used by Luginbuhl et al. through in situ enzymatic myristoylation of an ELP that contained a peptide substrate for myristoylation at the N‐terminus (Figure b) . They did so by a single step, one‐pot recombinant synthesis that used the transcription machinery of an E. coli reprogrammed to perform enzymatic lipidation within the cell.…”

“…These lipid‐ELP biopolymers differ from ELP diblocks and ELP‐peptide fusions because the micellar core of lipid‐ELPs contains lipids rather than only peptides, so it is significantly more dehydrated, allowing physical encapsulation of hydrophobic molecules in the core. Indeed, the ELP‐myristoyl micelle encapsulates hydrophobic chemotherapeutics such as paclitaxel and doxorubicin more efficiently than the ELP diblock micelle …”