RRC ID 37895
著者 Nagahama M, Seike S, Shirai H, Takagishi T, Kobayashi K, Takehara M, Sakurai J.
タイトル Role of P2X7 receptor in Clostridium perfringens beta-toxin-mediated cellular injury.
ジャーナル Biochim Biophys Acta
Abstract BACKGROUND:Clostridium perfringens beta-toxin is a pore-forming toxin (PFT) and an important agent of necrotic enteritis and enterotoxemia. We recently reported that beta-toxin strongly induced cell death in THP-1 cells via the formation of oligomers. We here describe that the P2X(7) receptor, which is an ATP receptor, interacts with beta-toxin.
METHODS:We tested the role of P2X(7) receptor in beta-toxin-induced toxicity using specific inhibitors, knockdown of receptor, expression of the receptor and interaction by dot-blot assay. The potency of P2X(7) receptor was further determined using an in vivo mouse model.
RESULTS:Selective P2X(7) receptor antagonists (oxidized ATP (o-ATP), oxidized ADP, and Brilliant Blue G (BBG)) inhibited beta-toxin-induced cytotoxicity in THP-1 cells. o-ATP also blocked the binding of beta-toxin to cells. The P2X(7) receptor and beta-toxin oligomer were localized in the lipid rafts of THP-1 cells. siRNA for the P2X(7) receptor inhibited toxin-induced cytotoxicity and binding of the toxin. In contrast, the siRNA knockdown of P2Y(2) or P2Y(6) had no effect on beta-toxin-induced cytotoxicity. The addition of beta-toxin to P2X(7)-transfected HEK-293 cells resulted in binding of beta-toxin oligomer. Moreover, beta-toxin specifically bound to immobilized P2X(7) receptors in vitro and colocalized with the P2X(7) receptor on the THP-1 cell surface. Furthermore, beta-toxin-induced lethality in mice was blocked by the preadministration of BBG.
CONCLUSIONS:The results of this study indicate that the P2X(7) receptor plays a role in beta-toxin-mediated cellular injury.
GENERAL SIGNIFICANCE:P2X(7) receptor is a potential target for the treatment of C. perfringens type C infection.
巻・号 1850(11)
ページ 2159-67
公開日 2015-11-1
DOI 10.1016/j.bbagen.2015.08.011
PII S0304-4165(15)00220-2
PMID 26299247
MeSH ADAM Proteins / physiology ADAM10 Protein Amyloid Precursor Protein Secretases / physiology Animals Bacterial Toxins / toxicity* Calcium / metabolism HEK293 Cells Humans Membrane Proteins / physiology Mice Mice, Inbred ICR RNA, Small Interfering / pharmacology Receptors, Purinergic P2X7 / physiology* Rosaniline Dyes / pharmacology
IF 3.411
引用数 17
WOS 分野 BIOPHYSICS BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
ヒト・動物細胞 THP-1(RCB1189) 293(RCB1637)