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| Proteins, Expression, Isolation and Analysis | … Cleared lysates were then bound to Ni-NTA resin (GenScript) or glutathione resin (GenScript) for 4 hours, with rolling at 4C. Beads were washed extensively before eluting for 1 hour in His elution buffer (PBS plus 500 mM imidazole, pH 7.4) or GST elution buffer (50 mM Tris-HCl… | Get A Quote |
Although serine metabolism plays a crucial role in the proliferation and survival of tumor cells, how it supports tumor cell migration remains poorly understood. Phosphoglycerate dehydrogenase (PHGDH) catalyzes the oxidation of 3-phosphoglycerate to 3-phosphonooxypyruvate, the first committed step in de novo serine biosynthesis. Here we show that PHGDH was monoubiquitinated by cullin 4A-based E3 ligase complex at lysine 146 in colorectal cancer (CRC) cells, which enhanced PHGDH activity by recruiting a chaperone protein, DnaJ homolog subfamily A member 1, to promote its tetrameric formation, thereby increasing the levels of serine, glycine, and S-adenosylmethionine (SAM). Increased levels of SAM upregulated the... More
