Mitochondrial ATP synthase is a target for TNBS-induced protein carbonylation in XS-106 dendritic cells.
ROS are produced in dendritic cells (DCs) during antigen presentation in contact hypersensitivity (CHS). As a result, ROS cause a number of nonenzymatic protein modifications, including carbonylation, which is the most widely used marker of oxidative stress. 2,4,6-Trinitrobenzene sulfonic acid (TNBS) is a well-characterized contact allergen that results in the formation of ROS. However, proteins that are carbonylated in DCs in response to TNBS have not been identified. To study ROS-dependent protein carbonylation in response to TNBS, we used the well-established mouse DC line, XS-106. We focused on the effects of TNBS on oxidation by examining selected oxidative markers. We identified TNBS-induced ROS and myeloperoxidase (MPO) proteins and demonstrated that the increase in ROS resulted in IL-12 production. The increase in oxidation was further confirmed by an oxidation-dependent increase in protein modifications, such as carbonylation. In fact, TNBS strongly induced carbonylation of mitochondrial adenosine triphosphate (ATP) synthase in XS-106 DCs, as determined by MALDI-TOF analysis and 2-D Western blotting. ROS production and protein carbonylation were confirmed in human monocyte-derived DCs (Mo-DCs). Furthermore, glutathione (GSH) decreased ROS and protein carbonylation in Mo-DCs. Carbonylation of ATP synthase in DCs may contribute to the pathophysiology of CHS.
Department of Dermatology and Cutaneous Biology Research Institute, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea.
Jeong Hwan
JH
Je
Tae Hyung
TH
Lee
Dong Hyun
DH
Kim
Young Hun
YH
Cho
Ju Hee
JH
Lee
Soo Chan
SC
Kim
Sang-Kyou
SK
Lee
Jaewon
J
Lee
Min-Geol
MG
Lee
D000818:Animals; D000831:Animals, Newborn; D018929:Cell Culture Techniques; D002460:Cell Line; D017077:Culture Media, Conditioned; D003713:Dendritic Cells / Q000187:drug effects* / Q000378:metabolism; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D051379:Mice; D025261:Mitochondrial Proton-Translocating ATPases / Q000378:metabolism*; D009000:Monocytes / Q000166:cytology; D009195:Peroxidase / Q000235:genetics / Q000378:metabolism; D050050:Protein Carbonylation / Q000187:drug effects*; D040901:Proteomics / Q000379:methods; D017382:Reactive Oxygen Species / Q000378:metabolism; D012867:Skin / Q000166:cytology; D014302:Trinitrobenzenesulfonic Acid / Q000494:pharmacology*
D017077:Culture Media, Conditioned; D017382:Reactive Oxygen Species; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D014302:Trinitrobenzenesulfonic Acid; D009195:Peroxidase; D025261:Mitochondrial Proton-Translocating ATPases