SIMPLE binds specifically to PI4P through SIMPLE-like domain and participates in protein trafficking in the trans-Golgi network and/or recycling endosomes
Fig 1
SIMPLE binds specifically to PI4P through the SIMPLE-like domain.
(A) Schematic representation of the indicated membrane lipids on a PIP strip. (B) MBP-SIMPLE and MBP alone, as a negative control, were incubated with a PIP strip and detected as described in the Materials and Methods. (C) Schematic representation of SIMPLE deletion constructs. Numbers indicate SIMPLE amino acid residues. SLD is the SIMPLE-like domain. (D) CBB-stained bands of purified MBP or MBP-tagged human-SIMPLE. (E) Alignment of 115–141 and the 115–141 anagram. Conserved cysteine residues are indicated in green. (F) Binding of MBP-fusion proteins (see D) to PIP strips: each protein (5 nM) was added to a PIP strip. (G) Binding of MBP-SIMPLE with cysteine-to-serine (CS) mutantion to PIP strips.