Extended Data Fig. 10: The binding-site mutations in MFSD1 lose binding to peptides in the thermal stability assay.
From: MFSD1 with its accessory subunit GLMP functions as a general dipeptide uniporter in lysosomes
(a) FSEC analysis of MFSD1WT and mutants normalized to the fluorescent signal at λex = 488 nm/λem = 510 nm of GFP (F488) of MFSD1WT. The supernatant of soluble fraction after whole-cell solubilization was loaded onto a Superose 6 5/150 column. (b) SDS-PAGE of purified MFSD1WT and binding site mutants. For each lane, 2 µg of protein were loaded. (c) Comparison of SEC traces of binding site mutants (colored) to MFSD1WT (grey) of each mutant. Each mutant was purified once for subsequent experiments. (d), (e) Melting temperatures derived from thermal stability experiments of each mutant in the absence (apo, grey) or presence of 5 mM of selected peptides. n = 3 independent experiments with data shown as mean ± SD. For mutants for which no bar graph is given, unfolding traces are given for the apo state to show that no TM value could be determined. Source numerical data and unprocessed blots are available in source data.
