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Proteins, Expression, Isolation and Analysis | … Supernatant was collected and the pellet was resuspended in 25 mM Tris-HCl (pH 80) Supernatant and pelleted fractions were analyzed with SDS-PAGE followed by eStain L1 (GenScript, China) staining The intensity of each band was quantified with ImageJ … | Get A Quote |
Intracellular compartmentation is a key strategy for the functioning of a cell. In 2010, several studies revealed that the metabolic enzyme CTP synthase (CTPS) can form filamentous structures termed cytoophidia in prokaryotic and eukaryotic cells. However, recent structural studies showed that CTPS only forms inactive product-bound filaments in bacteria while forming active substrate-bound filaments in eukaryotic cells. In this study, using negative staining and cryo-electron microscopy, we demonstrate that Drosophila CTPS, whether in substrate-bound or product-bound form, can form filaments. Our results challenge the previous model and indicate that substrate-bound and product-bound filaments can coexist in t... More