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Peroxiredoxin‐4 Interacts with Dopamine D5 Receptor to Reduce Endoplasmic Reticulum Stress
Author(s) -
Amatya Bibhas,
Yang Sufei,
Yu Peiying,
Armando Ines,
Zeng Chunyu,
Felder Robin A.,
Asico Laureano D.,
Jose Pedro A.,
Lee Hewang
Publication year - 2022
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2022.36.s1.r3865
Subject(s) - fenoldopam , endoplasmic reticulum , chemistry , hek 293 cells , oxidative stress , receptor , gene silencing , microbiology and biotechnology , unfolded protein response , agonist , biology , biochemistry , gene
Peroxiredoxin‐4 (PRDX4), an endoplasmic reticulum (ER)‐localized antioxidant enzyme, plays an essential role in cellular redox homeostasis by reducing hydrogen peroxide from thiol‐containing compounds. The dopamine D5 Receptor (D5R) also plays a protective role against oxidative stress. We hypothesize that D 5 R interacts with the PRDX4 to reduce oxidative stress in the kidney. In D 5 R‐HEK 293 cells, fenoldopam (FEN, 25 nM/12 hr, n=4), a D 1 R and D 5 R receptor agonist, increased PRDX4 protein expression (1.92±0.12‐fold over basal level, n=4), mainly in non‐lipid raft (LR) fractions (LRs: 24.9±11.4%, non‐LRs: 75.1±11.4%, baseline; LRs: 30.9±13.9%, non‐LRs: 174.1±16.7%, FEN). By contrast, fenoldopam did not affect PRDX4 protein expression in D 1 R‐HEK 293 cells, indicating a D 5 R‐specific effect on PRDX4 protein expression. In human renal proximal tubule cells (hRPTCs) and D 5 R HEK293 cells, fenoldopam increased the co‐immunoprecipitation between PRDX4 and D 5 R and their co‐localization in the ER. SiRNA‐mediated silencing of PRDX4 increased hydrogen peroxide production in both the vehicle (Veh)‐ and fenoldopam‐treated hRPTCs [(scrambled siRNA: 100 ±15.1% and 55.2± 7.2% with Veh and FEN, respectively; PRDX4 siRNA: 161.8±15.3% and 145.1±14.6 % with Veh and FEN, respectively, n=4)]. The D 5 R protects against inflammation and siRNA silencing of PRDX4 increased the production of the pro‐inflammatory cytokines, interleukin‐1β [26.88±3.8 and 46.40±4.2 pg/mL (n=3, D 5 R‐HEK 293); 15.87±1.2 and 37.9±1.4 pg/mL (n=3, hRPTCs), and tumor necrosis factors [131.7±6.5 and 271.2±18.1 pg/mL (n=4, D 5 R‐HEK 293); 108.8±11.8 and 240.1±13.7 pg/mL (n=4, hRPTCs)]. In D 5 R‐HEK293 and hRPTCs, the fenoldopam‐mediated decrease in ER‐resident caspase‐12 was also impaired by gene silencing of PRDX4 . Furthermore, PRDX4 protein expression was reduced in the kidney cortices of Drd5 ‐/‐ mice, relative to Drd5 WT mice (WT: 1.00±0.12, n=4; Drd5 ‐/‐ : 0.64±0.13, n=4; P<0.05). We conclude that D 5 R positively interacts with PRDX4 to reduce ER stress in the kidney.