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. 2020 May 13;16(5):e9370. doi: 10.15252/msb.20199370

Figure 3. Streptavidin depletion using prS beads allows for better enrichment and identification of biotinylated targets and their interactors.

Figure 3

  • A–D
    SEC61B BioID. (A) Relative intensity of streptavidin peptides in 100 ng or 1ug BioID experiments performed with either wtS or prS beads. The negative control only expressed BirA*. (B) Intensity‐based ranking of identified proteins in 1ug experiment. (C) Intensity of SEC61B and its top interacting proteins in 100 ng or 1ug BioID experiments using either wtS or prS beads. (D) Number of PSMs in BioID experiments.
  • E–H
    Integral membrane proteins. (E) Percentage of streptavidin peptides after enrichment of biotinylated membrane proteins, identified in 10% of wtS or prS sample, and in the residual 90% of the prS sample. (F) Intensity‐based ranking of proteins identified in the experiments in (E). (G) Number of total and membrane proteins identified in the experiments in (E). (H) Number of PSMs in the experiments in (E).
  • I–L
    Sphingolipid (SL)‐binding proteins. (I) Relative intensity of streptavidin after lipid enrichment in prS beads. (J) Intensity‐based ranking of proteins identified using either NeutrAvidin or prS beads. (K) Intensity of common sphingolipid‐binding proteins identified after enrichment via NeutrAvidin (NA, black) and prS beads (orange). (L) Number of PSMs in the respective experiments in (K).
Data information: Error bars in panels (A, C, D, I, K, and L) indicate standard deviation of three replicates.