Figure 1.

Schematic illustration of the TLR interaction proteomic approach. Murine endosomal TLR3, TLR7, TLR8, and TLR9 were stably expressed in the macrophage cell line RAW264.7 by retroviral transduction. After TAP of the receptor complexes, eluates were analyzed by one-dimensional SDS-PAGE and silver stained. 20 gel slices were excised from each lane, the proteins were digested, and the peptides were analyzed by LC-MSMS. All bait TLRs were recovered with the average amount of peptides and sequence coverage as indicated in the table. At ∼55 kD, several peptides were identified from CD14. A schematic illustration of CD14 is depicted with the domains shown (signal peptide in red and leucine-rich repeats in blue) and the location of the identified peptides (black bars). For each TLR, the average number of peptides (av p.) identified and the sequence coverage (av%sc) of CD14 are indicated in the table.