Basigin-MCT1 fusion constructs, expression in yeast, and transport functionality.
2021
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Zugriff:
( A ) Shown are fusion constructs of basigin (black/gray; PDB #6LZ0) with MCT1 (orange; PDB #6LZ0). Disulfide bridges are shown as orange spheres, and Glu218 of the basigin transmembrane helix is highlighted in red. ( B ) Western blot showing expression of MCT1 alone (58 kDa), and fused with BSGΔIg (WT, 67 kDa) or the E27R point mutant. Proteins were detected using a Penta-His antibody. ( C ) Live-cell confocal microscopy of fusion proteins via C-terminal GFP and soluble GFP (—). ( D, E ) Uptake of 14 C-labeled l-lactate into jen1Δ ady2Δ yeast at pH 6.8 and a 1 mM inward gradient. Shown are curves for non-expressing cells (background, D, ○), cells expressing MCT1 alone (D, ●), and in fusion with BSGΔIg (E, ■), or BSGΔIg E27R (E, □). The data are normalized to 1 mg of cells and the background of non-expressing cells was subtracted. Error bars indicate ± S.E.M. from three biological replicates.
Titel: |
Basigin-MCT1 fusion constructs, expression in yeast, and transport functionality.
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Autor/in / Beteiligte Person: | Anna-Lena Köpnick (10474898) ; Annika Jansen (10474901) ; Katharina Geistlinger (10126011) ; Nathan Hugo Epalle (10474904) ; Eric Beitz (147186) |
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Veröffentlichung: | 2021 |
Medientyp: | Bild |
DOI: | 10.1371/journal.pone.0249110.g001 |
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