Role of the Na(+)/K(+)-ATPase beta-subunit in peptide-mediated transdermal drug delivery.
In this work, we discovered that the Na(+)/K(+)-ATPase beta-subunit (ATP1B1) on epidermal cells plays a key role in the peptide-mediated transdermal delivery of macromolecular drugs. First, using a yeast two-hybrid assay, we screened candidate proteins that have specific affinity for the short peptide TD1 (ACSSSPSKHCG) identified in our previous work. Then, we verified the specific binding of TD1 to ATP1B1 in yeast and mammalian cells by a pull-down ELISA and an immunoprecipitation assay. Finally, we confirmed that TD1 mainly interacted with the C-terminus of ATP1B1. Our results showed that the interaction between TD1 and ATP1B1 affected not only the expression and localization of ATP1B1, but also the epidermal structure. In addition, this interaction could be antagonized by the exogenous competitor ATP1B1 or be inhibited by ouabain, which results in the decreased delivery of macromolecular drugs across the skin. The discovery of a critical role of ATP1B1 in the peptide-mediated transdermal drug delivery is of great significance for the future development of new transdermal peptide enhancers.
Changli
C
Wang
Renquan
R
Ruan
¶Department of Urology, Anhui Medical University, Hefei, Anhui 230032, China.
Li
L
Zhang
Yunjiao
Y
Zhang
Wei
W
Zhou
Jun
J
Lin
Weiping
W
Ding
Longping
L
Wen
D000279:Administration, Cutaneous; D000818:Animals; D045744:Cell Line, Tumor; D004337:Drug Carriers; D016503:Drug Delivery Systems; D004797:Enzyme-Linked Immunosorbent Assay; D005982:Glutathione Transferase / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D046911:Macromolecular Substances; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D010042:Ouabain / Q000737:chemistry; D010455:Peptides / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D051381:Rats; D017207:Rats, Sprague-Dawley; D012867:Skin / Q000187:drug effects*; D000254:Sodium-Potassium-Exchanging ATPase / Q000737:chemistry*; D020798:Two-Hybrid System Techniques
C503551:ATP1B1 protein, human; D004337:Drug Carriers; D046911:Macromolecular Substances; D010455:Peptides; D010042:Ouabain; D005982:Glutathione Transferase; D000254:Sodium-Potassium-Exchanging ATPase
K-ATPase beta-subunit; Na; macromolecular drug; peptide enhancer; transdermal drug delivery