Increasing the length and hydrophobicity of the C-terminal sequence of transthyretin strengthens its binding affinity to retinol binding protein.

Transthyretin (TTR) is a transporter for thyroid hormone (TH) and retinol, the latter via binding with retinol binding protein (RBP). Both the N-terminal and C-terminal regions of the TTR subunit are located in close proximity to the central binding channel for ligands. During the evolution of vertebrates, these regions changed in length and hydropathy. The changes in the N-terminal sequence were demonstrated to affect the binding affinities for THs and RBP. Here, the effects of changes in the C-terminal sequence were determined. Three chimeric TTRs, namely pigC/huTTR (human TTR with the C-terminal sequence changed to that of TTR), xenoN/pigC/huTTR (human TTR with the N-terminal and C-terminal sequences changed to those of and , respectively), and pigC/crocTTR ( TTR with the C-terminal sequence changed to that of TTR), were constructed and their binding affinities for human RBP were determined at low TTR/RBP molar ratio using chemiluminescence immunoblotting. The binding dissociation constant () values of pigC/huTTR, xenoN/pigC/huTTR and pigC/crocTTR were 3.20 ± 0.35, 1.53 ± 0.38 and 0.31 ± 0.04 μm, respectively, and the values of human and TTR were 4.92 ± 0.68 and 1.42 ± 0.45 μm, respectively. These results demonstrate chimeric TTRs bound RBP with a higher strength than wild-type TTRs, and the changes in the C-terminal sequence of TTR had a positive effect on its binding affinity for RBP. In addition, changes to the N-terminal and C-terminal sequences showed comparable effects on the binding affinity.