Blocking the Interactions between Calcium-Bound S100A12 Protein and the V Domain of RAGE Using Tranilast.
The receptor for advanced glycation end products (RAGE), a transmembrane receptor in the immunoglobulin superfamily, is involved in several inflammatory processes. RAGE induces cellular signaling pathways upon binding with various ligands, such as advanced glycation end products (AGEs), β-amyloids, and S100 proteins. The solution structure of S100A12 and the V ligand-binding region of RAGE have been reported previously. Using heteronuclear NMR spectroscopy to conduct 1H-15N heteronuclear single quantum coherence (HSQC) titration experiments, we identified and mapped the binding interface between S100A12 and the V domain of RAGE. The NMR chemical shift data were used as the constraints for the High Ambiguity Driven biomolecular DOCKing (HADDOCK) calculation to generate a structural model of the S100A12-V domain complex. In addition, tranilast (an anti-allergic drug) showed strong interaction with S100A12 in the 1H-15N HSQC titration, fluorescence experiments, and WST-1 assay. The results also indicated that tranilast was located at the binding site between S100A12 and the V domain, blocking interaction between these two proteins. Our results provide the mechanistic details for a structural model and reveal a potential precursor for an inhibitor for pro-inflammatory diseases, which could be useful for the development of new drugs.
Department of Chemistry, National Tsing Hua University, Hsinchu, 30013, Taiwan.
Jian Wei
JW
Chiou
Northwood High School, Irvine, California, 92620, United States of America.
Brian
B
Fu
Graduate Institute of Cancer Biology and Center for Molecular Medicine, China Medical University, Taichung, 40402, Taiwan.|Department of Biotechnology, Asia University, Taichung, 41354, Taiwan.
Ruey-Hwang
RH
Chou
Department of Chemistry, National Tsing Hua University, Hsinchu, 30013, Taiwan.
Chin
C
Yu
D020816:Amino Acid Motifs; D018926:Anti-Allergic Agents / Q000737:chemistry*; D001665:Binding Sites; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007700:Kinetics; D062105:Molecular Docking Simulation; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D000072756:Protein Conformation, alpha-Helical; D000072757:Protein Conformation, beta-Strand; D054730:Protein Interaction Domains and Motifs; D000067759:Receptor for Advanced Glycation End Products / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000067616:S100A12 Protein / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013816:Thermodynamics; D062367:ortho-Aminobenzoates / Q000737:chemistry*
C000593871:AGER protein, human; D018926:Anti-Allergic Agents; D000067759:Receptor for Advanced Glycation End Products; D011994:Recombinant Proteins; D000067616:S100A12 Protein; C090441:S100A12 protein, human; D062367:ortho-Aminobenzoates; C012293:tranilast