Fig. 2: Validation of MP3-seq with coiled-coil heterodimers.
From: Massively parallel measurement of protein–protein interactions by sequencing using MP3-seq
a, NICP series 1×1s13 with complementary heptads designed to interact shown in like colors. b, MP3-seq LFC of the NICP series interactions. All MP3-seq values were calculated from five biological replicates except for those labeled, where labels indicate the number of replicates available. Yellow outlines denote designed on-target interactions. c, Correlation of the on-target and off-target NICP series MP3-seq LFCs with average fold activation fluorescence values13 (n = 141 PPIs; three homodimers had insufficient reads or were autotuned and were omitted). The gray bar is the gap separating on-target and off-target interactions. LFCs are presented as the LFC ± standard error (SE). d,e, Filtering NICP series interactions (d) and P, PA and N series designed coil interactions47 (e) to include only those with Padj ≤ 0.05. Line weights correspond to MP3-seq P-LFCs. f, A designed 1×1 and its truncations from a previous study48. g, MP3-seq enrichment for the AN and BN coils and their truncations from three biological replicates. A gray ‘×’ indicates a missing interaction. h, Correlation of n = 9 AN and BN truncation PPI MP3-seq P-LFCs with Kd values from Thomas et al.48. P-LFCs are presented as the P-LFC ± SE and dissociation constants are presented as the mean ± s.d.
