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Clarified supernatants were filtered using 0.2-m filters, transferred to new 50-ml Falcon tubes containing Ni-NTA resin equilibrated with lysis buffer (20 mM Tris-HCl (pH 9), 10 mM imidazole, 200 mM NaCl and 5% glycerol) and agitated for 1 h to allow protein binding

By contrast, serine itself is an allosteric inhibitor of PHGDH, and this allosteric inhibition can spontaneously disappear at low serine levels (Sugimoto and Pizer, 1968)

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