Fig. 3: Mvp1 Mut1 is dimeric and is defective in lipid binding and CPY sorting in vivo. | Nature Communications

Fig. 3: Mvp1 Mut1 is dimeric and is defective in lipid binding and CPY sorting in vivo.

From: The cryo-EM structure of the SNX–BAR Mvp1 tetramer

Fig. 3

The absolute molecular weights of Mvp1 Mut1 (a) or Mvp1 K198A (purple) or Mvp1 R199A, I200A (pink) (b) were determined using SEC-MALS as in Fig. 1. Proteins were eluted in buffer containing 250 mM NaCl. c Subcellular localization of EGFP-tagged Mvp1 and its mutants in Δmvp1 cells. Vacuoles were stained with the lipophilic dye FM 4–64. Images were obtained by confocal microscopy. Scale bar—5 μm. d CPY secretion assays. W303A or Δmvp1 cells expressing vector or the indicated construct were plated onto SD-Trp to maintain plasmid selection (left) or YPD (right) plates and incubated at 30 °C for 24 h. Plates were then overlaid with nitrocellulose for an additional 16 h. CPY secretion was detected by immunoblotting using an anti-CPY antibody. The leftmost spot in each case is 2 μl of a OD600 = 0.5 culture: spots to the right of this are sequential fivefold dilutions. For d source data are provided as a Source Data file.

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