title: Expression, purification and transduction of PEP-1-botulinum neurotoxin type A (PEP-1-BoNT/A) into skin.
abstract: Botulinum neurotoxin A (BoNT/A) has been used therapeutically to treat muscular hypercontractions and sudomotor hyperactivity and it has been reported that BoNT/A might have analgesic properties in headache. PEP-1 peptide is a known carrier peptide that delivers full-length native proteins in vitro and in vivo. In this study, a BoNT/A gene were fused with PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame PEP-1-BoNT/A fusion protein. The expressed and purified PEP-1-BoNT/A fusion proteins were efficiently transduced into cells in a time- and dose-dependent manner when added exogenously in a culture medium. In addition, immunohistochemical analysis revealed that PEP-1-BoNT/A fusion protein efficiently penetrated into the epidermis as well as the dermis of the subcutaneous layer, when sprayed on mice skin. These results suggest that PEP-1-BoNT/A fusion protein provide an efficient strategy for therapeutic delivery in various human diseases related to this protein.
authors.affiliation: Department of Biomedical Sciences and Research Institute for Bioscience and Biotechnology, Hallym University, Korea.
authors.forename: Dae Won
authors.initials: DW
authors.lastname: Kim
authors.forename: So Young
authors.initials: SY
authors.lastname: Kim
authors.forename: Jae Jin
authors.initials: JJ
authors.lastname: An
authors.forename: Sun Hwa
authors.initials: SH
authors.lastname: Lee
authors.forename: Sang Ho
authors.initials: SH
authors.lastname: Jang
authors.forename: Moo Ho
authors.initials: MH
authors.lastname: Won
authors.forename: Tae Cheon
authors.initials: TC
authors.lastname: Kang
authors.forename: Kwang-Hoe
authors.initials: KH
authors.lastname: Chung
authors.forename: Hyun-Ho
authors.initials: HH
authors.lastname: Jung
authors.forename: Sung-Woo
authors.initials: SW
authors.lastname: Cho
authors.forename: Jin Hi
authors.initials: JH
authors.lastname: Choi
authors.forename: Jinseu
authors.initials: J
authors.lastname: Park
authors.forename: Won Sik
authors.initials: WS
authors.lastname: Eum
authors.forename: Soo Young
authors.initials: SY
authors.lastname: Choi
mesh_terms: D000287:Administration, Topical; D000818:Animals; D015153:Blotting, Western; D019274:Botulinum Toxins, Type A / Q000008:administration & dosage* / Q000235:genetics / Q000493:pharmacokinetics; D003543:Cysteamine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000032:analysis / Q000493:pharmacokinetics; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D010455:Peptides / Q000008:administration & dosage* / Q000032:analysis / Q000493:pharmacokinetics; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000235:genetics / Q000493:pharmacokinetics; D012867:Skin / Q000737:chemistry / Q000378:metabolism*
chemical_list: C503924:Pep-1 peptide; D010455:Peptides; D011993:Recombinant Fusion Proteins; D003543:Cysteamine; D019274:Botulinum Toxins, Type A; D016650:Fluorescein-5-isothiocyanate