{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T08:30:37Z","timestamp":1789115437977,"version":"build-2803163510"},"reference-count":41,"publisher":"Public Library of Science (PLoS)","issue":"4","license":[{"start":{"date-parts":[[2024,4,16]],"date-time":"2024-04-16T00:00:00Z","timestamp":1713225600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000165","name":"Division of Chemistry","doi-asserted-by":"publisher","award":["CHE2145906"],"award-info":[{"award-number":["CHE2145906"]}],"id":[{"id":"10.13039\/100000165","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100001159","name":"Hawaii Community Foundation","doi-asserted-by":"publisher","award":["HCF40846 (013357-00002)"],"award-info":[{"award-number":["HCF40846 (013357-00002)"]}],"id":[{"id":"10.13039\/100001159","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000009","name":"Foundation for the National Institutes of Health","doi-asserted-by":"publisher","award":["R35GM127303"],"award-info":[{"award-number":["R35GM127303"]}],"id":[{"id":"10.13039\/100000009","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>The chemokines of the immune system act as first responders by operating as chemoattractants, directing immune cells to specific locations of inflamed tissues. This promiscuous network is comprised of 50 ligands and 18 receptors where the ligands may interact with the receptors in various oligomeric states i.e., monomers, homodimers, and heterodimers. Chemokine receptors are G-protein coupled receptors (GPCRs) present in the membrane of immune cells. The migration of immune cells occurs in response to a concentration gradient of the ligands. Chemotaxis of neutrophils is directed by CXC-ligand (CXCL) activation of the membrane bound CXC chemokine receptor 2 (CXCR2). CXCR2 plays an important role in human health and is linked to disorders such as autoimmune disorders, inflammation, and cancer. Yet, despite their important role, little is known about the biophysical characteristics controlling ligand:ligand and ligand:receptor interaction essential for biological activity. In this work, we study the homodimers of three of the CXCR2 cognate ligands, CXCL1, CXCL5, and CXCL8. The ligands share high structural integrity but a low sequence identity. We show that the sequence diversity has evolved different binding affinities and stabilities for the CXC-ligands resulting in diverse agonist\/antagonist behavior. Furthermore, CXC-ligands fold through a three-state mechanism, populating a folded monomeric state before associating into an active dimer.<\/jats:p>","DOI":"10.1371\/journal.pone.0298418","type":"journal-article","created":{"date-parts":[[2024,4,16]],"date-time":"2024-04-16T17:23:07Z","timestamp":1713288187000},"page":"e0298418","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":5,"title":["Biophysical characterization of the CXC chemokine receptor 2 ligands"],"prefix":"10.1371","volume":"19","author":[{"given":"Patrick","family":"Martin","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emily A.","family":"Kurth","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"Budean","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nathalie","family":"Momplaisir","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elaine","family":"Qu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4591-3036","authenticated-orcid":true,"given":"Jennifer M.","family":"Simien","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Grace E.","family":"Orellana","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chad A.","family":"Brautigam","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alan V.","family":"Smrcka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6629-2793","authenticated-orcid":true,"given":"Ellinor","family":"Haglund","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2024,4,16]]},"reference":[{"issue":"2","key":"pone.0298418.ref001","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1016\/S1074-7613(00)80165-X","article-title":"Chemokines: a new classification system and their role in immunity","volume":"12","author":"A Zlotnik","year":"2000","journal-title":"Immunity"},{"issue":"4","key":"pone.0298418.ref002","doi-asserted-by":"crossref","first-page":"570","DOI":"10.1016\/j.cell.2017.04.004","article-title":"Metabolic Instruction of Immunity","volume":"169","author":"MD Buck","year":"2017","journal-title":"Cell"},{"key":"pone.0298418.ref003","doi-asserted-by":"crossref","first-page":"609","DOI":"10.1146\/annurev-immunol-032713-120236","article-title":"Metabolic regulation of immune responses","volume":"32","author":"K Ganeshan","year":"2014","journal-title":"Annu Rev Immunol"},{"issue":"9","key":"pone.0298418.ref004","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1038\/nri.2016.70","article-title":"A guide to immunometabolism for immunologists","volume":"16","author":"LA O\u2019Neill","year":"2016","journal-title":"Nat Rev Immunol"},{"issue":"16","key":"pone.0298418.ref005","doi-asserted-by":"crossref","first-page":"2944","DOI":"10.1111\/febs.14466","article-title":"A guide to chemokines and their receptors","volume":"285","author":"CE Hughes","year":"2018","journal-title":"FEBS J"},{"issue":"4","key":"pone.0298418.ref006","doi-asserted-by":"crossref","DOI":"10.3390\/ijms23042168","article-title":"CXCR2 Receptor: Regulation of Expression, Signal Transduction, and Involvement in Cancer","volume":"23","author":"J Korbecki","year":"2022","journal-title":"Int J Mol Sci"},{"issue":"5","key":"pone.0298418.ref007","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1007\/s10540-006-9025-9","article-title":"Structural basis of chemokine receptor function\u2014a model for binding affinity and ligand selectivity","volume":"26","author":"L Rajagopalan","year":"2006","journal-title":"Biosci Rep"},{"issue":"11","key":"pone.0298418.ref008","doi-asserted-by":"crossref","first-page":"4083","DOI":"10.1073\/pnas.0508863103","article-title":"Identification of the minimal protein-folding nucleus through loop-entropy perturbations","volume":"103","author":"MO Lindberg","year":"2006","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"41","key":"pone.0298418.ref009","doi-asserted-by":"crossref","first-page":"27904","DOI":"10.1074\/jbc.M801776200","article-title":"Changes of protein folding pathways by circular permutation. 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Kauzmann","year":"1959","journal-title":"Adv Protein Chem"},{"issue":"2","key":"pone.0298418.ref033","doi-asserted-by":"crossref","first-page":"660","DOI":"10.1006\/jmbi.1993.1417","article-title":"Contribution of hydration to protein folding thermodynamics. II. The entropy and Gibbs energy of hydration","volume":"232","author":"PL Privalov","year":"1993","journal-title":"J Mol Biol"},{"issue":"3","key":"pone.0298418.ref034","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1016\/S0022-2836(03)00627-2","article-title":"Comparison of the folding processes of distantly related proteins. 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