Fructose-6-phosphate 2-kinase and fructose 2,6-bisphosphatase have been partially purified from maize leaves by PEG fractionadon and by chromatography on TSK-DEAE ion exchanger and Blue-Sepharose 4B. Fructose-6-phosph...Fructose-6-phosphate 2-kinase and fructose 2,6-bisphosphatase have been partially purified from maize leaves by PEG fractionadon and by chromatography on TSK-DEAE ion exchanger and Blue-Sepharose 4B. Fructose-6-phosphate 2-kinase was activated by phosphate and inhibited by 3-pnosphoglycerate. Furctose 2,6-bisphosphatase was inhibited by inorganic phosphate and fructose-6-phosphate.The observed pattern of reguladon suggests that systhesis and degradation of Fru-2,6-P<sub>2</sub> respond to changes in the concentration of effectors. An increase in the level of glycerate-3-phosphate or dihydroxyacetonephosphate will result in a decrease in the level of Fru-2,6-P<sub>2</sub>. Conversely a rise in Fru-6-P concentration will lead to an increase in the Fru-2, 6-P<sub>2</sub> concentration.展开更多
To study the roles and association of murine double minute gene 2(MDM2)and fructose-1,6-biphosphatase(FBP1)in human hepatocellular carcinoma(HCC),growth response of human HCC cells was assessed using proliferation and...To study the roles and association of murine double minute gene 2(MDM2)and fructose-1,6-biphosphatase(FBP1)in human hepatocellular carcinoma(HCC),growth response of human HCC cells was assessed using proliferation and apoptosis assay.Pro-survival AKT signaling associated proteins(p-AKT,survivin and cleaved caspase 3)were assessed using western blotting.The correlation between MDM2 and FBP1 was assessed using coimmunoprecipitation combined with ubiquitination assay.Our data suggested that low expression of FBP1 was correlated with high levels of MDM2 in HCC cell lines(Huh7 and Hep3B).Overexpression of FBP1 resulted in antiproliferation,pro-apoptosis,the up-regulation of cleaved caspase 3 while the downregulation of survivin and phosphor(p)-AKT,however,knockdown of FBP1 led to the opposite.Furthermore,overexpression of MDM2 potently reversed FBP1-induced proliferation inhibition and apoptosis,while Nutlin-3(an MDM2 inhibitor)reversed the change trends induced by FBP1 knockdown in the aforementioned events.Lastly,but not least importantly,our data elucidated that MDM2 binds directly to FBP1 and promotes FBP1 ubiquitination.In conclusion,our data firstly suggested the involvement of FBP1 and its association with MDM2 in HCC cell growth.MDM2-FBP1-regulated HCC cell growth and the activation of AKT were mediated,at least in part,through FBP1 degradation.展开更多
文摘Fructose-6-phosphate 2-kinase and fructose 2,6-bisphosphatase have been partially purified from maize leaves by PEG fractionadon and by chromatography on TSK-DEAE ion exchanger and Blue-Sepharose 4B. Fructose-6-phosphate 2-kinase was activated by phosphate and inhibited by 3-pnosphoglycerate. Furctose 2,6-bisphosphatase was inhibited by inorganic phosphate and fructose-6-phosphate.The observed pattern of reguladon suggests that systhesis and degradation of Fru-2,6-P<sub>2</sub> respond to changes in the concentration of effectors. An increase in the level of glycerate-3-phosphate or dihydroxyacetonephosphate will result in a decrease in the level of Fru-2,6-P<sub>2</sub>. Conversely a rise in Fru-6-P concentration will lead to an increase in the Fru-2, 6-P<sub>2</sub> concentration.
基金The study was supported by the Grant QN201706 from Young Talent Training Project of Health Development Planning Commission of Changzhou City,and Grant CE20205034 from Science and Technology Supporting Project of Changzhou City.
文摘To study the roles and association of murine double minute gene 2(MDM2)and fructose-1,6-biphosphatase(FBP1)in human hepatocellular carcinoma(HCC),growth response of human HCC cells was assessed using proliferation and apoptosis assay.Pro-survival AKT signaling associated proteins(p-AKT,survivin and cleaved caspase 3)were assessed using western blotting.The correlation between MDM2 and FBP1 was assessed using coimmunoprecipitation combined with ubiquitination assay.Our data suggested that low expression of FBP1 was correlated with high levels of MDM2 in HCC cell lines(Huh7 and Hep3B).Overexpression of FBP1 resulted in antiproliferation,pro-apoptosis,the up-regulation of cleaved caspase 3 while the downregulation of survivin and phosphor(p)-AKT,however,knockdown of FBP1 led to the opposite.Furthermore,overexpression of MDM2 potently reversed FBP1-induced proliferation inhibition and apoptosis,while Nutlin-3(an MDM2 inhibitor)reversed the change trends induced by FBP1 knockdown in the aforementioned events.Lastly,but not least importantly,our data elucidated that MDM2 binds directly to FBP1 and promotes FBP1 ubiquitination.In conclusion,our data firstly suggested the involvement of FBP1 and its association with MDM2 in HCC cell growth.MDM2-FBP1-regulated HCC cell growth and the activation of AKT were mediated,at least in part,through FBP1 degradation.
