{"doi":"10.1007/978-1-0716-2593-4_32","title":"Melatonin Binding to Human NQO2 by Isothermal Titration Calorimetry","abstract":null,"journal":"Methods in Molecular Biology","year":2022,"id":686305,"datarank":0.26876392038420827,"base_score":1.791759469228055,"endowment":1.791759469228055,"self_citation_contribution":0.26876392038420827,"citation_network_contribution":0.0,"self_endowment_contribution":0.26876392038420827,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":5,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1508743,"name":"Gilles Ferry","orcid":null,"position":1,"is_corresponding":false},{"id":118450,"name":"Jean A. Boutin","orcid":null,"position":2,"is_corresponding":false},{"id":141762,"name":"Barbara Calamini","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Melatonin Binding to Human NQO2 by Isothermal Titration Calorimetry","abstract":"To ensure the physical interaction between a protein and its ligand, many techniques can be applied. One of them, isothermal titration calorimetry (ITC), measures the heat exchange between a forming molecular complex and its milieu. From this heat exchange, it is possible to acquire the thermodynamic parameters, the binding stoichiometry and the affinity constant (K<sub>a</sub>) between the two interacting binding partners, which can then be used to determine the dissociation constant (K<sub>d</sub>). We made use of ITC to determine the true K<sub>d</sub> of melatonin for its putative receptor MT3, also known as the enzyme quinone reductase 2 (NQO2). In this chapter, we describe the step-by-step procedure for performing this experiment and extend it to 2-iodomelatonin, a melatonin derivative that was used in the initial identification and characterization of MT3. The dissociation constants of melatonin and 2-iodomelatonin toward NQO2 derived from these experiments are in line with data reported previously, albeit using alternative techniques.","is_dataset_classified":null,"base_score":1.791759469228055,"endowment":1.791759469228055,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"36180701","pmcid":null,"openalex_id":"https://openalex.org/W4298326575","authors":[],"funders":[],"total_grants":0,"fwci":12.8265,"citation_percentile":0.98805732,"influential_citations":0,"citation_trend":[{"year":2022,"count":1},{"year":2023,"count":1},{"year":2024,"count":1},{"year":2025,"count":1},{"year":2026,"count":1}],"oa_status":"closed","license":"https://www.springer.com/tdm","oa_locations":[{"url":"https://link.springer.com/content/pdf/10.1007/978-1-0716-2593-4_32","host_type":"publisher"},{"url":"https://doi.org/10.1007/978-1-0716-2593-4_32","host_type":"book series"},{"url":"https://pubmed.ncbi.nlm.nih.gov/36180701","host_type":"repository"},{"url":"https://ut3-toulouseinp.hal.science/hal-04056225","host_type":"repository"}],"fields_of_study":["thermodynamics and calorimetric analyses","Circadian rhythm and melatonin","Free Radicals and Antioxidants","Calorimetry","Humans","Ligands","Melatonin","Protein Binding","Quinone Reductases","Thermodynamics"],"mesh_terms":["Calorimetry","Humans","Ligands","Melatonin","Protein Binding","Quinone Reductases","Thermodynamics"],"keywords":["Isothermal titration calorimetry","Dissociation constant","Melatonin","Chemistry","Binding constant","Titration","Calorimetry","Dissociation (chemistry)","Isothermal process","Allosteric regulation","Stoichiometry","Biophysics","Binding site","Physical chemistry","Thermodynamics","Enzyme","Biochemistry","Receptor","Biology","Endocrinology","Ligand binding","Association constant","Quinone Reductase 2","Binding Stoichiometry","2-Iodo-melatonin","Enthalpy And Entropy Of Binding"],"sdg_mappings":[{"sdg_number":0,"sdg_label":"Clean water and sanitation"}],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-18T19:08:35.725339Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}