Selection of Amino Acids on TLR2 for Mutant Development

As negatively charged pectin (DM7) could inhibit TLR2, we analyzed amino acids lying close to the ligand binding site on TLR2. We chose R315, R316, R321, and K347 amino acids (R-arginine and K-lysine) as putative pectin binding site on TLR2. R321 and R347 were selected as these positively charged amino acids were involved in binding between TLR2 and TLR1 interfaces (35). R315 and R316 were selected as these amino acids were structurally close to the ligand binding site of TLR2. The structural proximity of these amino acids on TLR2 structure was confirmed using PyMOL program (Schrödinger, New York, NY, USA).