Fig. 4
From: Characterizing glucokinase variant mechanisms using a multiplexed abundance assay
A conformational shift towards the super-open state as a mechanism for variants in the 150–179 region. A Left, protein structures of wild-type glucokinase in the closed and super-open states with the 150–179 region marked in orange. Right, overview of glucokinase in the super-open state with the 150–179 region substituted by 30 (15xGS) or 6 residues of GS (3xGS) shown in red. B Yeast growth assay scoring the activity of wild-type glucokinase (WT) and the two mutants. The growth on galactose is used as a control while growth on 0.2% glucose reflects glucokinase activity. C Western blot showing the protein levels of the indicated constructs expressed in the hxk1\(\Delta\) hxk2\(\Delta\) glk1\(\Delta\) yeast strain from panel B. D DHFR-PCA probing the abundance of wild-type glucokinase (WT) and the two mutants by growing yeast cells on control medium (DMSO) and medium with methotrexate (MTX) to select for abundance. E Structure of glucokinase in the closed state with the 150–179 region marked in black, E157 in dark blue and K161 in red. F Yeast growth assay scoring the activity of wild-type glucokinase (WT) and the indicated single and double mutants. PDBs: 1V4S (closed) and 1V4T (super-open)