tetano
Editor, Senior Moderator
J Biochem
. 2023 Mar 8;mvad020.
doi: 10.1093/jb/mvad020. Online ahead of print.
Reactive oxygen species are associated with the inhibitory effect of N-(4-hydroxyphenyl)-retinamide on the entry of the severe acute respiratory syndrome-coronavirus 2
Yasuhiro Hayashi[SUP] 1 [/SUP], Xuhao Huang[SUP] 2 [/SUP], Takashi Tanikawa[SUP] 3 [/SUP], Kazunari Tanigawa[SUP] 4 [/SUP], Mizuki Yamamoto[SUP] 5 [/SUP], Jin Gohda[SUP] 5 [/SUP], Jun-Ichiro Inoue[SUP] 6 [/SUP], Koichi Fukase[SUP] 2 7 [/SUP], Kazuya Kabayama[SUP] 2 7 [/SUP]
Affiliations
Abstract
N-(4-hydroxyphenyl)-retinamide (4-HPR) inhibits the dihydroceramide Δ4-desaturase 1 (DEGS1) enzymatic activity. We previously reported that 4-HPR suppresses the severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) spike protein-mediated membrane fusion through a decrease in membrane fluidity in a DEGS1-independent manner. However, the precise mechanism underlying the inhibition of viral entry by 4-HPR remains unclear. In this study, we examined the role of reactive oxygen species (ROS) in the inhibition of membrane fusion by 4-HPR, since 4-HPR is a well-known ROS-inducing agent. Intracellular ROS generation was found to be increased in the target cells in a cell-cell fusion assay following 4-HPR treatment, which was attenuated by the addition of the antioxidant, α-tocopherol (TCP). The reduction in membrane fusion susceptibility by 4-HPR treatment in the cell-cell fusion assay was alleviated by TCP addition. Furthermore, fluorescence recovery after photobleaching analysis showed that the lateral diffusion of glycosylphosphatidylinositol-anchored protein and SARS CoV-2 receptor was reduced by 4-HPR treatment and restored by TCP addition. These results indicate that the decrease in SARS-CoV-2 spike protein-mediated membrane fusion and membrane fluidity by 4-HPR was due to ROS generation. Taken together, these results demonstrate that ROS production is associated with the 4-HPR inhibitory effect on SARS-CoV-2 entry.
Keywords: 4-HPR; ROS; SARS-CoV-2; TCP.
. 2023 Mar 8;mvad020.
doi: 10.1093/jb/mvad020. Online ahead of print.
Reactive oxygen species are associated with the inhibitory effect of N-(4-hydroxyphenyl)-retinamide on the entry of the severe acute respiratory syndrome-coronavirus 2
Yasuhiro Hayashi[SUP] 1 [/SUP], Xuhao Huang[SUP] 2 [/SUP], Takashi Tanikawa[SUP] 3 [/SUP], Kazunari Tanigawa[SUP] 4 [/SUP], Mizuki Yamamoto[SUP] 5 [/SUP], Jin Gohda[SUP] 5 [/SUP], Jun-Ichiro Inoue[SUP] 6 [/SUP], Koichi Fukase[SUP] 2 7 [/SUP], Kazuya Kabayama[SUP] 2 7 [/SUP]
Affiliations
- PMID: 36888972
- DOI: 10.1093/jb/mvad020
Abstract
N-(4-hydroxyphenyl)-retinamide (4-HPR) inhibits the dihydroceramide Δ4-desaturase 1 (DEGS1) enzymatic activity. We previously reported that 4-HPR suppresses the severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) spike protein-mediated membrane fusion through a decrease in membrane fluidity in a DEGS1-independent manner. However, the precise mechanism underlying the inhibition of viral entry by 4-HPR remains unclear. In this study, we examined the role of reactive oxygen species (ROS) in the inhibition of membrane fusion by 4-HPR, since 4-HPR is a well-known ROS-inducing agent. Intracellular ROS generation was found to be increased in the target cells in a cell-cell fusion assay following 4-HPR treatment, which was attenuated by the addition of the antioxidant, α-tocopherol (TCP). The reduction in membrane fusion susceptibility by 4-HPR treatment in the cell-cell fusion assay was alleviated by TCP addition. Furthermore, fluorescence recovery after photobleaching analysis showed that the lateral diffusion of glycosylphosphatidylinositol-anchored protein and SARS CoV-2 receptor was reduced by 4-HPR treatment and restored by TCP addition. These results indicate that the decrease in SARS-CoV-2 spike protein-mediated membrane fusion and membrane fluidity by 4-HPR was due to ROS generation. Taken together, these results demonstrate that ROS production is associated with the 4-HPR inhibitory effect on SARS-CoV-2 entry.
Keywords: 4-HPR; ROS; SARS-CoV-2; TCP.