title: Self-assembling amphipathic alpha-helical peptides induce the formation of active protein aggregates in vivo.
abstract: We recently found that several self-assembling alpha, beta, or surfactant-like peptides, when terminally attached to proteins, can promote the in vivo assembly of active protein aggregates (or active inclusion bodies, AIBs) in Escherichia coil. In this work, we systematically examined the AIBs induced by an amphipathic alpha-helical peptide 18Awt (EWLKAFYEKVLEKLKELF) and its variants with altered ion pairs. Transmission electron microscopic and Fourier transform infrared spectroscopic analyses suggested that the AIBs appeared to adopt an amorphous mesh-like structure, and were likely induced by helical structures formed by the assembly of the 18A peptides. Confocal fluorescent micrographic analysis revealed that the AIBs resided around the periphery of the cell membrane or in the cytoplasm, depending on the distribution of ion pairs on the 18A peptides, which suggested that the association between the aggregates and the cell membrane was mediated by the lipid-18A interaction. Two of these 18A peptide variants were further used in constructing cleavable self-aggregating tags (cSAT) in conjunction with an intein molecule for protein purification, and verified using two model proteins. This extends the cSAT approach for laboratory and potentially industrial uses. Our study might also provide new insights into aggregation-related diseases.
authors.forename: Zhanglin
authors.initials: Z
authors.lastname: Lin
authors.forename: Bihong
authors.initials: B
authors.lastname: Zhou
authors.forename: Wei
authors.initials: W
authors.lastname: Wu
authors.forename: Lei
authors.initials: L
authors.lastname: Xing
authors.forename: Qing
authors.initials: Q
authors.lastname: Zhao
mesh_terms: D000595:Amino Acid Sequence; D046529:Microscopy, Electron, Transmission; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D010455:Peptides; D011506:Proteins