The molecular chaperone HSP60 is a chaperonin homolog of GroEL. We had previously shown that the immunosuppressant mizoribine is bound directly to HSP60 and inhibited its chaperone activity. However, the inhibitory me...The molecular chaperone HSP60 is a chaperonin homolog of GroEL. We had previously shown that the immunosuppressant mizoribine is bound directly to HSP60 and inhibited its chaperone activity. However, the inhibitory mechanisms of HSP60 by mizoribine have not yet been fully understood. In the present study, we investigated the influence of mizoribine on a folding cycle of HSP60 and co-chaperone HSP10. Our results showed that mizoribine inhibited the folding cycle of HSP60/HSP10. The ATPase activity of HSP60/HSP10 was decreased in the presence of mizoribine and the dissociation of HSP10 from HSP-60 was also decreased by mizoribine. The same functions of GroEL and/or GroES were slightly affected by mizoribine. Based on our findings, we discuss the inhibitory mechanisms of HSP60 by mizoribine.展开更多
Insecticidal effects of 4-hexylresorcinol, a phenoloxidase （PO） inhibitor, were determined on Hyphantria cunea （Drury） under laboratory conditions. The LC50 for the 15- d-old larvae was estimated to be 2.95 g/L af...Insecticidal effects of 4-hexylresorcinol, a phenoloxidase （PO） inhibitor, were determined on Hyphantria cunea （Drury） under laboratory conditions. The LC50 for the 15- d-old larvae was estimated to be 2.95 g/L after 96 h exposure. The activities of glutathione S-transferase （GST） and PO showed a decrease in larvae treated with 4-hexylresorcinol, and the IC50 of GST and PO were estimated to be 0.8 and 0.43 g/L, respectively, 24 h after treatment. The PO from the hemolymph of fall webworm was purified by ammonium sulfate precipitation, gel-filtration, and ion-exchange chromatography, and then enzymatic characteristics and the mechanism of inhibition were determined using L-dihydroxyphenylalanine （L-DOPA） as the substrate. The purified PO showed a single band on SDS-PAGE with a molecular weight of about 70 kDa. The optimum pH for PO activity was observed at pH 7.0, optimum temperature was found to be 45 ℃, and PO activity was strongly inhibited by Zn2＋. IC50 values were estimated to be 8.2, 19.14, and 24.04/zmol/L for 4-hexylresorsinol, kojic acid, and quercetin, respectively. The inhibitory potencies （i.e., 150 of each compound/Is0 of 4-hexylresorcinol） ofkojic acid and quercetin on H. cunea PO were estimated to be 1.87 and 2.89, respectively. 4-hexylresorcinol was determined to be a competitive inhibitor, and kojic acid and quercetin were determined to be mixed inhibitors. PO is one of the most important enzymes in an insect＇s immune system, and the use ofPO inhibitors seems to be a promising approach for pest control due to their potential safety for humans.展开更多
文摘The molecular chaperone HSP60 is a chaperonin homolog of GroEL. We had previously shown that the immunosuppressant mizoribine is bound directly to HSP60 and inhibited its chaperone activity. However, the inhibitory mechanisms of HSP60 by mizoribine have not yet been fully understood. In the present study, we investigated the influence of mizoribine on a folding cycle of HSP60 and co-chaperone HSP10. Our results showed that mizoribine inhibited the folding cycle of HSP60/HSP10. The ATPase activity of HSP60/HSP10 was decreased in the presence of mizoribine and the dissociation of HSP10 from HSP-60 was also decreased by mizoribine. The same functions of GroEL and/or GroES were slightly affected by mizoribine. Based on our findings, we discuss the inhibitory mechanisms of HSP60 by mizoribine.
文摘Insecticidal effects of 4-hexylresorcinol, a phenoloxidase （PO） inhibitor, were determined on Hyphantria cunea （Drury） under laboratory conditions. The LC50 for the 15- d-old larvae was estimated to be 2.95 g/L after 96 h exposure. The activities of glutathione S-transferase （GST） and PO showed a decrease in larvae treated with 4-hexylresorcinol, and the IC50 of GST and PO were estimated to be 0.8 and 0.43 g/L, respectively, 24 h after treatment. The PO from the hemolymph of fall webworm was purified by ammonium sulfate precipitation, gel-filtration, and ion-exchange chromatography, and then enzymatic characteristics and the mechanism of inhibition were determined using L-dihydroxyphenylalanine （L-DOPA） as the substrate. The purified PO showed a single band on SDS-PAGE with a molecular weight of about 70 kDa. The optimum pH for PO activity was observed at pH 7.0, optimum temperature was found to be 45 ℃, and PO activity was strongly inhibited by Zn2＋. IC50 values were estimated to be 8.2, 19.14, and 24.04/zmol/L for 4-hexylresorsinol, kojic acid, and quercetin, respectively. The inhibitory potencies （i.e., 150 of each compound/Is0 of 4-hexylresorcinol） ofkojic acid and quercetin on H. cunea PO were estimated to be 1.87 and 2.89, respectively. 4-hexylresorcinol was determined to be a competitive inhibitor, and kojic acid and quercetin were determined to be mixed inhibitors. PO is one of the most important enzymes in an insect＇s immune system, and the use ofPO inhibitors seems to be a promising approach for pest control due to their potential safety for humans.
