Pentamidine blocks the interaction between mutant S100A5 and RAGE V domain and inhibits the RAGE signaling pathway.
The human S100 protein family contains small, dimeric and acidic proteins that contain two EF-hand motifs and bind calcium. When S100A5 binds calcium, its conformation changes and promotes interaction with the target protein. The extracellular domain of RAGE (Receptor of Advanced Glycation End products) contain three domains: C1, C2 and V. The RAGE V domain is the target protein of S100A5 that promotes cell survival, growth and differentiation by activating several signaling pathways. Pentamidine is an apoptotic and antiparasitic drug that is used to treat or prevent pneumonia. Here, we found that pentamidine interacts with S100A5 using HSQC titration. We elucidated the interactions of S100A5 with RAGE V domain and pentamidine using fluorescence and NMR spectroscopy. We generated two binary models-the S100A5-RAGE V domain and S100A5-Pentamidine complex-and then observed that the pentamidine and RAGE V domain share a similar binding region in mS100A5. We also used the WST-1 assay to investigate the bioactivity of S100A5, RAGE V domain and pentamidine. These results indicated that pentamidine blocks the binding between S100A5 and RAGE V domain. This finding is useful for the development of new anti-proliferation drugs.
Department of Chemistry, National Tsing Hua University, No. 101, Section 2, Kuang-Fu Road, Hsinchu 30013, Taiwan. Electronic address: ccjwo@yahoo.com.tw.
Ching Chang
CC
Cho
Graduate Institute of Cancer Biology and Center for Molecular Medicine, China Medical University, No.91, Hsueh-Shih Road, Taichung 40402, Taiwan. Electronic address: rhchou@mail.cmu.edu.tw.
Ruey Hwang
RH
Chou
Department of Chemistry, National Tsing Hua University, No. 101, Section 2, Kuang-Fu Road, Hsinchu 30013, Taiwan. Electronic address: cyu.nthu@gmail.com.
Chin
C
Yu
D000951:Antigens, Neoplasm / Q000737:chemistry* / Q000378:metabolism*; D001665:Binding Sites; D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D020928:Mitogen-Activated Protein Kinases / Q000737:chemistry* / Q000378:metabolism*; D008956:Models, Chemical; D062105:Molecular Docking Simulation; D009154:Mutation; D010419:Pentamidine / Q000008:administration & dosage* / Q000737:chemistry*; D011485:Protein Binding; D000072417:Protein Domains; D009418:S100 Proteins / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D015398:Signal Transduction / Q000187:drug effects
D000951:Antigens, Neoplasm; D009418:S100 Proteins; C082355:S100A5 protein, human; D010419:Pentamidine; C498978:MOK protein, human; D020928:Mitogen-Activated Protein Kinases
HADDOCK; NMR solution structure; Protein–ligand interaction; RAGE; S100 protein; WST-1 assay