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KMID : 0380220060390050642
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Journal of Biochemistry and Molecular Biology
2006 Volume.39 No. 5 p.642 ~ p.647
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Expression, Purification and Transduction of PEP-1-Botulinum Neurotoxin Type A (PEP-1-BoNT/A) into Skin
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Kim Dae-Won
Kim So-Young
An Jae-Jin
Lee Sun-Hwa
Jang Sang-Ho
Woo Moo-Ho
Kang Tae-Cheon
Chung Kwang-Hoe
Jung Hyun-Ho
Cho Sung-Woo
Park Jin-Seu
Eum Won-Sik
Choi Soo-Young
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Abstract
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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 fulllength 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.
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KEYWORD
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Botulinum toxin A (BoNT/A), Facial wrinkles, PEP-1 peptide, Protein therapy, Protein transduction
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