title: The protein truncation caused by fusion of PEP-1 peptide and protective roles of transduced PEP-1-MsrA in skin cells.
abstract: PEP-1 peptide has been used for transduction of native protein into mammalian cells. This work describes the findings that the fusion of PEP-1 to target proteins led to protein truncation likely in a non-protein-specific manner. Approximately 75% of PEP-1-MsrA fusion protein was truncated in the N-terminal region of MsrA between Lys-27 and Val-28 during expression in Escherichia coli and purification. This large protein truncation was also observed in another PEP-1 fused protein, PEP-1- MsrB2, in the N-terminal region of MsrB2. The full-length PEP-1-MsrA protein was rapidly transduced into keratinocyte cells within 15 min. The transduced PEP-1-MsrA was functionally active and could protect skin cells against oxidative stress-and ultraviolet radiation-induced cell death. Collectively, our data demonstrated the protective roles of MsrA in skin cells and, moreover, may raise a concern of protein truncation caused by fusion of PEP-1 about the general use of this peptide for protein transduction.
authors.affiliation: Department of Biochemistry and Molecular Biology, Yeungnam University College of Medicine, Daegu 705-717, Korea.
authors.forename: Tae-Hyung
authors.initials: TH
authors.lastname: Lee
authors.forename: Seung Hee
authors.initials: SH
authors.lastname: Choi
authors.forename: Hwa-Young
authors.initials: HY
authors.lastname: Kim
mesh_terms: D017209:Apoptosis; D002460:Cell Line; D003543:Cysteamine / Q000031:analogs & derivatives* / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism; D051150:Methionine Sulfoxide Reductases / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D018384:Oxidative Stress; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D055550:Protein Stability; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014466:Ultraviolet Rays
chemical_list: C503924:Pep-1 peptide; D010455:Peptides; D011993:Recombinant Fusion Proteins; D003543:Cysteamine; D051150:Methionine Sulfoxide Reductases; C027219:methionine sulfoxide reductase