Figure 3.

Representation of amino acids in the membrane-binding region of D4 analyzed with the combinatorial library.A, occurrence of sequences in the combinatorial library and after selection against lipid vesicles with different lipid compositions as indicated. B–D, relative abundance of amino acid residues at randomized positions after selection against vesicles containing only POPC (B), POPC and 50 mol% Chol (C), and POPC and 50 mol% acetyl-Chol (D). The color scale represents log₂ of amino acid residue frequencies normalized to the input combinatorial library. The amino acid residues in the boxes outlined in red represent WT PFO residues. E, sequence logos for all variants with alternative CRM. F, enrichment of nonsynonymous mutations observed at the indicated positions in the SSL and the combinatorial library. The sequence logos were generated with WebLogo 3 (62).