Skip to main content
. Author manuscript; available in PMC: 2021 Feb 1.
Published in final edited form as: FEBS Lett. 2019 Nov 16;594(4):646–664. doi: 10.1002/1873-3468.13648

Figure 9. HPLC Size Exclusion Chromatography of PKM2 and PKM2 Mutants.

Figure 9.

PKM2 proteins were prepared using Prep 3, then separated by HPLC size exclusion in the absence of FBP to determine the effect of mutations on tetramer-monomer equilibrium. This separation method accommodates a high concentration of enzyme (0.5 mg/ml) relative to the kinetic assays. An elution volume of ~8.25 mL is consistent with tetramers while an elution volume of ~9.25 mL is consistent with monomer. Only the G415R and R399E mutants showed significant amounts of protein eluting at a volume consistent with the enzyme being a dimer or monomer.