A New Nuclear Function of the Entamoeba histolytica Glycolytic Enzyme Enolase: The Metabolic Regulation of Cytosine-5 Methyltransferase 2 (Dnmt2) Activity (original) (raw)
Figure 3
In vivo interaction of Ehmeth with enolase.
Immunoprecipitation with an anti-HA antibody from a nuclear lysate of E. histolytica trophozoites that express Ehmeth as a CHH-tagged protein (pJST4-Ehmeth) grown in regular media (control) and from trophozoites grown in a glucose starvation media (glucose starvation). Detection of immunoprecipitated proteins was done by western blot with an anti-enolase antibody. To validate that the same amounts of Ehmeth were used in the assay, immunoprecipated proteins were analyzed with an anti His antibody which detects the CHH tagged Ehmeth. As a negative control, immunoprecipitation with an anti-HA antibody from a nuclear lysate of E. histolytica trophozoites that express CHH-klp5 was used (right panel). The physical interaction between enolase and Ehmeth is demonstrated only after immunoprecipitation from Ehmeth tagged trophozoites and this complex is enhanced following glucose starvation (3 fold according to Tina densitometry analysis).