title: Biophysical characterization of Ca2+-binding of S100A5 and Ca2+-induced interaction with RAGE.
abstract: S100A5 is a calcium-binding protein of S100 family, which represents a major ligand to the receptor for advanced glycation end product (RAGE), a pattern recognition receptor engaged in diverse pathological processes. Here we have characterized calcium binding of S100A5 and the complex formation between S100A5 and RAGE using calorimetry and NMR spectroscopy. S100A5 binds to calcium ions in a sequential manner with the equilibrium dissociation constants (KD) of 1.3 μM and 3.5 μM, which corresponds to the calcium-binding at the C-terminal and N-terminal EF-hands. Upon calcium binding, S100A5 interacts with the V domain of RAGE (RAGE-v) to form a heterotrimer (KD∼5.9 μM) that is distinct among the S100 family proteins. Chemical shift perturbation data from NMR titration experiments indicates that S100A5 employs the periphery of the dimer interface to interact with RAGE-v. Distinct binding mode and stoichiometry of RAGE against different S100 family proteins could be important to modulate diverse RAGE signaling.
authors.affiliation: Department of Agricultural Biotechnology, Seoul National University, 599 Gwanak-ro, Gwanak-gu, Seoul 151-921, South Korea. Electronic address: iktaekim@snu.ac.kr.
authors.forename: Iktae
authors.initials: I
authors.lastname: Kim
authors.affiliation: Department of Agricultural Biotechnology, Seoul National University, 599 Gwanak-ro, Gwanak-gu, Seoul 151-921, South Korea. Electronic address: koonlee@snu.ac.kr.
authors.forename: Ko On
authors.initials: KO
authors.lastname: Lee
authors.affiliation: Department of Agricultural Biotechnology, Seoul National University, 599 Gwanak-ro, Gwanak-gu, Seoul 151-921, South Korea.
authors.forename: Young-Joo
authors.initials: YJ
authors.lastname: Yun
authors.affiliation: Department of Agricultural Biotechnology, Seoul National University, 599 Gwanak-ro, Gwanak-gu, Seoul 151-921, South Korea.
authors.forename: Jea Yeon
authors.initials: JY
authors.lastname: Jeong
authors.affiliation: Division of Magnetic Resonance, Korea Basic Science Institute, 16 Yeongudanji-Ro, Ochang, Chungbuk 363-883, South Korea.
authors.forename: Eun-Hee
authors.initials: EH
authors.lastname: Kim
authors.affiliation: Division of Magnetic Resonance, Korea Basic Science Institute, 16 Yeongudanji-Ro, Ochang, Chungbuk 363-883, South Korea.
authors.forename: Haekap
authors.initials: H
authors.lastname: Cheong
authors.affiliation: Division of Magnetic Resonance, Korea Basic Science Institute, 16 Yeongudanji-Ro, Ochang, Chungbuk 363-883, South Korea.
authors.forename: Kyoung-Seok
authors.initials: KS
authors.lastname: Ryu
authors.affiliation: Advanced Analysis Center, Korea Institute of Science and Technology, Seoul 02792, South Korea.
authors.forename: Nak-Kyoon
authors.initials: NK
authors.lastname: Kim
authors.affiliation: Department of Agricultural Biotechnology, Seoul National University, 599 Gwanak-ro, Gwanak-gu, Seoul 151-921, South Korea. Electronic address: jysuh@snu.ac.kr.
authors.forename: Jeong-Yong
authors.initials: JY
authors.lastname: Suh
mesh_terms: D000951:Antigens, Neoplasm / Q000378:metabolism*; D002118:Calcium / Q000737:chemistry*; D002151:Calorimetry; D002845:Chromatography; D020832:EF Hand Motifs; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007700:Kinetics; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D011485:Protein Binding; D000072417:Protein Domains; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D009418:S100 Proteins / Q000378:metabolism*; D015398:Signal Transduction; D013816:Thermodynamics
chemical_list: D000951:Antigens, Neoplasm; D008024:Ligands; D009418:S100 Proteins; C082355:S100A5 protein, human; C498978:MOK protein, human; D020928:Mitogen-Activated Protein Kinases; D002118:Calcium
keywords: Ca(2+) binding; Calorimetry; NMR spectroscopy; RAGE; S100A5