Fig 3.

TIFA exists as an intrinsic dimer in solution. (A and B) Size exclusion chromatography (HiLoad 16/60 Superdex 75 pg column) (A) and AUC analysis (B) show that recombinant TIFA and mutants exist as dimers in solution. (C) Recombinant WT TIFA appeared as a dimer from low (0.5 mg/ml) to high (2 mg/ml) protein concentrations in size exclusion chromatography (Superdex 75 10/300 GL column). (D) AUC analysis of WT recombinant TIFA alone (I) or with the T9-containing peptide (II) or the pT9-containing peptide (III). The concentration of WT TIFA used was 10 mM, and the peptides were presented at a molar ratio of 1:10 (protein/peptide). (E) TIFA remained in the monomeric form on SDS-PAGE under a nonreduced condition, demonstrating that TIFA did not dimerize through disulfide bonds.