{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:48:17Z","timestamp":1787028497209,"version":"build-2736575974"},"reference-count":50,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2023,1,12]],"date-time":"2023-01-12T00:00:00Z","timestamp":1673481600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004281","name":"National Science Centre, Poland","doi-asserted-by":"publisher","award":["2013\/11\/B\/T5\/02271"],"award-info":[{"award-number":["2013\/11\/B\/T5\/02271"]}],"id":[{"id":"10.13039\/501100004281","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Free drug concentration in the blood sera is crucial for its appropriate activity. Serum albumin, the universal blood carrier protein, is responsible for transporting drugs and releasing them into the bloodstream. Therefore, a drug\u2019s binding to SA is especially important for its bioavailability and it is a key problem in the drug design process. In this paper, we present crystal structures of three animal serum albumin complexes: ovine, caprine, and leporine, with diclofenac, a popular non-steroidal anti-inflammatory drug that is used in therapy of chronic and acute pain. Details of diclofenac binding mode by the presented serum albumins are compared with analogous complexes of human and equine serum albumins. The analysis of the occupied binding pockets in crystal structures of the investigated serum albumins from different mammals shows that they have two common and a number of unique diclofenac binding sites. The most intriguing is the fact that the albumins from the described species are able to bind different numbers of molecules of this popular anti-inflammatory drug, but none of the binding sites overlap with ones in the human serum albumin.<\/jats:p>","DOI":"10.3390\/ijms24021534","type":"journal-article","created":{"date-parts":[[2023,1,13]],"date-time":"2023-01-13T01:30:26Z","timestamp":1673573426000},"page":"1534","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Structural Investigation of Diclofenac Binding to Ovine, Caprine, and Leporine Serum Albumins"],"prefix":"10.3390","volume":"24","author":[{"given":"Julita A.","family":"Talaj","sequence":"first","affiliation":[{"name":"Department of Molecular Biology of Cancer, Medical University of Lodz, 6\/8 Mazowiecka Street, 92-215 Lodz, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kamil","family":"Zielinski","sequence":"additional","affiliation":[{"name":"Department of Hematology, Comprehensive Cancer Center and Traumatology, Copernicus Memorial Hospital, Pabianicka Street 62, 93-513 Lodz, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0808-1349","authenticated-orcid":false,"given":"Anna","family":"Bujacz","sequence":"additional","affiliation":[{"name":"Institute of Molecular and Industrial Biotechnology, Lodz University of Technology, B. Stefanowskiego 2\/22, 90-537 Lodz, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,1,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Li, M., Jiang, M., Gao, Y., Zheng, Y., Liu, Z., and Zhou, C. (2016). Systems Pharmacology in Small Molecular Drug Discovery. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17020246"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1007\/s40262-013-0042-0","article-title":"Prediction of Drug Disposition on the Basis of Its Chemical Structure","volume":"52","author":"Stepensky","year":"2013","journal-title":"Clin. Pharmacokinet."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Peters, T. (1996). All About Albumin: Biochemistry, Genetics, and Medical Applications, Academic Press.","DOI":"10.1016\/B978-012552110-9\/50006-4"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1016\/j.mam.2011.12.002","article-title":"Human Serum Albumin: From Bench to Bedside","volume":"33","author":"Fanali","year":"2012","journal-title":"Mol. Asp. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"721","DOI":"10.1006\/jmbi.2000.4158","article-title":"Crystallographic Analysis Reveals Common Modes of Binding of Medium and Long-Chain Fatty Acids to Human Serum Albumin","volume":"303","author":"Bhattacharya","year":"2000","journal-title":"J. Mol. Biol."},{"key":"ref_6","first-page":"1052","article-title":"Further Characterization of Specific Drug Binding Sites on Human Serum Albumin","volume":"12","author":"Sudlow","year":"1976","journal-title":"Mol. Pharmacol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1016\/S0022-2860(02)00256-9","article-title":"Interaction of Drugs with Bovine and Human Serum Albumin","volume":"614","author":"Sulkowska","year":"2002","journal-title":"J. Mol. Struct."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1016\/j.jmb.2005.07.075","article-title":"Structural Basis of the Drug-Binding Specificity of Human Serum Albumin","volume":"353","author":"Ghuman","year":"2005","journal-title":"J. Mol. Biol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"342","DOI":"10.2133\/dmpk.24.342","article-title":"Lessons from the Crystallographic Analysis of Small Molecule Binding to Human Serum Albumin","volume":"24","author":"Curry","year":"2009","journal-title":"Drug Metab. Pharmacokinet."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1002\/bdd.1829","article-title":"A Diclofenac Suppository\u2013nabumetone Combination Therapy for Arthritic Pain Relief and a Monitoring Method for the Diclofenac Binding Capacity of HSA Site II in Rheumatoid Arthritis","volume":"34","author":"Setoguchi","year":"2013","journal-title":"Biopharm. Drug Dispos."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1002\/bdd.1887","article-title":"Albumin-Binding of Diclofenac and the Effect of a Site II Inhibitor in the Aqueous Humor of Cataract Patients with the Instillation of Diclofenac","volume":"35","author":"Osaki","year":"2014","journal-title":"Biopharm. Drug Dispos."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"227","DOI":"10.15171\/apb.2016.031","article-title":"Interactions between Sirolimus and Anti-Inflammatory Drugs: Competitive Binding for Human Serum Albumin","volume":"6","author":"Khodaei","year":"2016","journal-title":"Adv. Pharm. Bull."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"589","DOI":"10.15171\/apb.2016.073","article-title":"Characterization of the Effect of Drug-Drug Interaction on Protein Binding in Concurrent Administration of Sulfamethoxazol and Diclofenac Sodium Using Bovine Serum Albumin","volume":"6","author":"Hossain","year":"2016","journal-title":"Adv. Pharm. Bull."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1088","DOI":"10.1016\/j.neuropharm.2005.07.004","article-title":"Non-Steroidal Anti-Inflammatory Drugs Have Anti-Amyloidogenic Effects for Alzheimer\u2019s \u03b2-Amyloid Fibrils in Vitro","volume":"49","author":"Hirohata","year":"2005","journal-title":"Neuropharmacology"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"639","DOI":"10.1016\/j.neurobiolaging.2006.03.013","article-title":"McGeer. NSAIDs and Alzheimer Disease: Epidemiological, Animal Model and Clinical Studies","volume":"28","author":"McGeer","year":"2007","journal-title":"Neurobiol. Aging"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"543","DOI":"10.1016\/S0006-2952(97)00342-0","article-title":"Biochemical Pharmacology of Nonsteroidal Anti-Inflammatory Drugs","volume":"55","author":"Wu","year":"1998","journal-title":"Biochem. Pharmacol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"410","DOI":"10.1107\/S0907444906003660","article-title":"Specific Binding of Non-Steroidal Anti-Inflammatory Drugs (NSAIDs) to Phospholipase A2: Structure of the Complex Formed between Phospholipase A2 and Diclofenac at 2.7 Resolution","volume":"62","author":"Singh","year":"2006","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"19","DOI":"10.3121\/cmr.2007.698","article-title":"An Evidence-Based Update on Nonsteroidal Anti-Inflammatory Drugs","volume":"5","author":"Ong","year":"2007","journal-title":"Clin. Med. Res."},{"key":"ref_19","first-page":"81","article-title":"Evolution of Nonsteroidal Anti-Inflammatory Cyclooxygenase (COX) Inhibition and Beyond Drugs (NSAIDs)","volume":"11","author":"Rao","year":"2008","journal-title":"J. Pharm. Pharmaceut. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1016\/j.jct.2016.08.020","article-title":"Binding thermodynamics of Diclofenac and Naproxen with human and bovine serum albumins: A calorimetric and spectroscopic study","volume":"103","author":"Sprague","year":"2016","journal-title":"J. Chem. Thermodyn."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1016\/j.ejps.2012.03.007","article-title":"Insights into the Binding of the Drugs Diclofenac Sodium and Cefotaxime Sodium to Serum Albumin: Calorimetry and Spectroscopy","volume":"46","author":"Sharma","year":"2012","journal-title":"Eur. J. Pharm. Sci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1016\/j.ejps.2014.04.002","article-title":"Calorimetric Investigation of Diclofenac Drug Binding to a Panel of Moderately Glycated Serum Albumins","volume":"59","author":"Indurthi","year":"2014","journal-title":"Eur. J. Pharm. Sci."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"945","DOI":"10.1002\/bio.3055","article-title":"Urea-Induced Binding between Diclofenac Sodium and Bovine Serum Albumin: A Spectroscopic Insight","volume":"31","author":"Dohare","year":"2016","journal-title":"Luminescence"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"19","DOI":"10.4172\/2254-609X.100033","article-title":"Studies on Binding of Widely used Drugs with Human Serum Albumin at Different Temperatures and PHs","volume":"5","author":"Yasseen","year":"2016","journal-title":"J. Biomed. Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1016\/j.saa.2013.01.093","article-title":"Investigations on the Interactions of Diclofenac Sodium with HSA and ctDNA Using Molecular Modeling and Multispectroscopic Methods","volume":"110","author":"Cui","year":"2013","journal-title":"Spectrochim. Acta Part A Mol. Biomol. Spectrosc."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2077","DOI":"10.1080\/10826070802225338","article-title":"Estimation of Binding Constants for Diclofenac Sodium and Bovine Serum Albumin by Affinity Capillary Electrophoresis and Fluorescence Spectroscopy","volume":"31","author":"Wang","year":"2008","journal-title":"J. Liq. Chromatogr. Relat. Technol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1021\/acs.jmedchem.5b00909","article-title":"Structural Insights into the Competitive Binding of Diclofenac and Naproxen by Equine Serum Albumin","volume":"59","author":"Sekula","year":"2016","journal-title":"J. Med. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1178","DOI":"10.1111\/cbdd.12583","article-title":"Structural Basis of Non-Steroidal Anti-Inflammatory Drug Diclofenac Binding to Human Serum Albumin","volume":"86","author":"Zhang","year":"2015","journal-title":"Chem. Biol. Drug Des."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"630","DOI":"10.1038\/nature02317","article-title":"Diclofenac Residues as the Cause of Vulture Population Decline in Pakistan","volume":"427","author":"Oaks","year":"2004","journal-title":"Nature"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1098\/rsbl.2005.0425","article-title":"Toxicity of Diclofenac to Gyps Vultures","volume":"2","author":"Swan","year":"2006","journal-title":"Biol. Lett."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"20130574","DOI":"10.1098\/rstb.2013.0574","article-title":"Avian Scavengers and the Threat from Veterinary Pharmaceuticals","volume":"369","author":"Cuthbert","year":"2014","journal-title":"Philos. Trans. R. Soc. B Biol. Sci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"16789","DOI":"10.1073\/pnas.0407607101","article-title":"Halogen Bonds in Biological Molecules","volume":"101","author":"Auffinger","year":"2004","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"5061","DOI":"10.1021\/jm100112j","article-title":"A Medicinal Chemist\u2019s Guide to Molecular Interactions","volume":"53","author":"Bissantz","year":"2010","journal-title":"J. Med. Chem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1278","DOI":"10.1107\/S0907444912027047","article-title":"Structures of Bovine, Equine and Leporine Serum Albumin","volume":"68","author":"Bujacz","year":"2012","journal-title":"Acta Crystallogr. Sect. D Biol. Crystallogr."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"442","DOI":"10.1107\/S0909049512006395","article-title":"Facilities for Macromolecular Crystallography at the Helmholtz-Zentrum Berlin","volume":"19","author":"Mueller","year":"2012","journal-title":"J. Synchrotron Rad."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1140\/epjp\/i2015-15141-2","article-title":"The macromolecular crystallography beamlines at BESSY II of the Helmholtz-Zentrum Berlin: Current status and perspectives","volume":"130","author":"Mueller","year":"2015","journal-title":"Eur. Phys. J. Plus"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"244","DOI":"10.1107\/S0021889806004717","article-title":"Effects of Cryoprotectant Concentration and Cooling Rate on Vitrification of Aqueous Solutions","volume":"39","author":"Berejnov","year":"2006","journal-title":"J. Appl. Crystallogr."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"789","DOI":"10.1107\/S0907444910015416","article-title":"Cryoprotection Properties of Salts of Organic Acids: A Case Study for a Tetragonal Crystal of HEW Lysozyme","volume":"66","author":"Bujacz","year":"2010","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1107\/S0907444909047337","article-title":"XDS","volume":"66","author":"Kabsch","year":"2010","journal-title":"Acta Crystallogr. D Biol. Cryst."},{"key":"ref_40","first-page":"133","article-title":"Integration, scaling, space-group assignment and post refinement","volume":"66","author":"Kabsch","year":"2010","journal-title":"Acta Crystallogr."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"240","DOI":"10.1107\/S0907444996012255","article-title":"Refinement of Macromolecular Structures by the Maximum-Likelihood Method","volume":"53","author":"Murshudov","year":"1997","journal-title":"Acta Crystallogr. D Biol Crystallogr."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"658","DOI":"10.1107\/S0021889807021206","article-title":"Phaser Crystallographic Software","volume":"40","author":"McCoy","year":"2007","journal-title":"J Appl. Crystallogr."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"896","DOI":"10.1107\/S205979831701470X","article-title":"Crystal Structures of Serum Albumins from Domesticated Ruminants and Their Complexes with 3,5-Diiodosalicylic Acid","volume":"73","author":"Bujacz","year":"2017","journal-title":"Acta Crystallogr. D Struct. Biol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1107\/S0907444911001314","article-title":"REFMAC5 for the Refinement of Macromolecular Crystal Structures","volume":"67","author":"Murshudov","year":"2011","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1107\/S0907444910045749","article-title":"Overview of the CCP4 Suite and Current Developments","volume":"67","author":"Winn","year":"2011","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"2126","DOI":"10.1107\/S0907444904019158","article-title":"Coot: Model-Building Tools for Molecular Graphics","volume":"60","author":"Emsley","year":"2004","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1107\/S0907444910007493","article-title":"Features and development of Coot","volume":"66","author":"Emsley","year":"2010","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1107\/S2059798316018210","article-title":"Polder Maps: Improving OMIT Maps by Excluding Bulk-Solvent","volume":"73","author":"Liebschner","year":"2017","journal-title":"Acta Crystallogr. D Struct. Biol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1107\/S0907444909042073","article-title":"MolProbity: All-Atom Structure Validation for Macromolecular Crystallography","volume":"66","author":"Chen","year":"2010","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"334","DOI":"10.1002\/chir.23162","article-title":"Structural investigations of stereoselective profen binding by equine and leporine serum albumins","volume":"32","author":"Zielinski","year":"2020","journal-title":"Chirality"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/2\/1534\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:04:32Z","timestamp":1760119472000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/2\/1534"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,12]]},"references-count":50,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2023,1]]}},"alternative-id":["ijms24021534"],"URL":"https:\/\/doi.org\/10.3390\/ijms24021534","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,1,12]]}}}