Expression, purification and transduction of PEP-1-botulinum neurotoxin type A (PEP-1-BoNT/A) into skin.
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.
Kim
Dae Won
DW
Department of Biomedical Sciences and Research Institute for Bioscience and Biotechnology, Hallym University, Korea.
Kim
So Young
SY
An
Jae Jin
JJ
Lee
Sun Hwa
SH
Jang
Sang Ho
SH
Won
Moo Ho
MH
Kang
Tae Cheon
TC
Chung
Kwang-Hoe
KH
Jung
Hyun-Ho
HH
Cho
Sung-Woo
SW
Choi
Jin Hi
JH
Park
Jinseu
J
Eum
Won Sik
WS
Choi
Soo Young
SY
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*
C503924:Pep-1 peptide; D010455:Peptides; D011993:Recombinant Fusion Proteins; D003543:Cysteamine; D019274:Botulinum Toxins, Type A; D016650:Fluorescein-5-isothiocyanate