Supplemental Material for Scharff-Poulsen, Moriya, and Johnston, 2018

Supplementary FIG Y1: Sequence alignment<br>Sequences of rat GLUT5, human GLUT1, human GLUT3, <i>Escherichia coli</i> XylE, and Staphylococcus epidermis GlcP, for which the 3D structures are known, are aligned with <i>Saccharomyces cerevisiae </i>Hxt1 and Rgt2. Numbering of amino acids is shown to the left of the sequences. Strictly conserved residues are highlighted in red-filled boxes, and highly conserved residues are shown in yellow-filled boxes. Boxes above the alignment indicate transmembrane helices (TM1 through TM12) and intracellular helices (ICH1 through ICH5) in rGLUT5. The substitutions that confer constitutive signaling of Rgt2 are indicated by arrows below the alignment.<br> <br>Supplementary FIG Y2: Structural alignment<br>Structural alignment of Bovine GLUT5 (4YB9) and <i>E. coli</i> XylE (4QIQ) in inward-open conformations. The GLUT5 (violet) and XylE (yellow) structures were aligned using PyMOL and are shown in a side-view. Visible transmembrane helices (TM) and intracellular helices (ICH) are indicated. The alignment shows that the two structures are very similar to each other.<br> <br>Supplementary FIG Y3: Imaging of live <i>S. cerevisiae </i>cells expressing GFP-fusions of Hxt1 wild type (upper panel) and of Hxt1-D82V (lower panel) from the GAL1 promoter. Cells were grown and induced with galactose as described in (Scharff-Poulsen and Pedersen 2013). Comparison of GFP fluorescence and phase contrast images show that the GFP fusions are highly expressed and accumulate in the plasma membrane of the cells.<br>