{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,25]],"date-time":"2026-07-25T02:07:18Z","timestamp":1784945238204,"version":"3.55.0"},"reference-count":29,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2021,6,17]],"date-time":"2021-06-17T00:00:00Z","timestamp":1623888000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000054","name":"National Cancer Institute","doi-asserted-by":"publisher","award":["F30 CA236182"],"award-info":[{"award-number":["F30 CA236182"]}],"id":[{"id":"10.13039\/100000054","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R37 AI058072"],"award-info":[{"award-number":["R37 AI058072"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"name":"National Institute for Health","award":["K08 DE026189"],"award-info":[{"award-number":["K08 DE026189"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Candida species cause serious infections requiring prolonged and sometimes toxic therapy. Antimicrobial proteins, such as chemokines, hold great interest as potential additions to the small number of available antifungal drugs. Metamorphic proteins reversibly switch between multiple different folded structures. XCL1 is a metamorphic, antimicrobial chemokine that interconverts between the conserved chemokine fold (an \u03b1\u2013\u03b2 monomer) and an alternate fold (an all-\u03b2 dimer). Previous work has shown that human XCL1 kills C. albicans but has not assessed whether one or both XCL1 folds perform this activity. Here, we use structurally locked engineered XCL1 variants and Candida killing assays, adenylate kinase release assays, and propidium iodide uptake assays to demonstrate that both XCL1 folds kill Candida, but they do so via different mechanisms. Our results suggest that the alternate fold kills via membrane disruption, consistent with previous work, and the chemokine fold does not. XCL1 fold-switching thus provides a mechanism to regulate the XCL1 mode of antifungal killing, which could protect surrounding tissue from damage associated with fungal membrane disruption and could allow XCL1 to overcome candidal resistance by switching folds. This work provides inspiration for the future design of switchable, multifunctional antifungal therapeutics.<\/jats:p>","DOI":"10.3390\/pathogens10060762","type":"journal-article","created":{"date-parts":[[2021,6,17]],"date-time":"2021-06-17T04:15:46Z","timestamp":1623903346000},"page":"762","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Metamorphic Protein Folding Encodes Multiple Anti-Candida Mechanisms in XCL1"],"prefix":"10.3390","volume":"10","author":[{"given":"Acacia F.","family":"Dishman","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI 53226, USA"},{"name":"Medical Scientist Training Program, Medical College of Wisconsin, Milwaukee, WI 53226, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"He","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Medical College of Wisconsin, Milwaukee, WI 53226, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brian F.","family":"Volkman","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI 53226, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna R.","family":"Huppler","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Medical College of Wisconsin, Milwaukee, WI 53226, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,6,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"ofy187","DOI":"10.1093\/ofid\/ofy187","article-title":"Epidemiology and Clinical Features of Invasive Fungal Infection in a US Health Care Network","volume":"5","author":"Webb","year":"2018","journal-title":"Open Forum Infect. Dis."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1097\/INF.0000000000002530","article-title":"Etiology and Outcome of Candidemia in Neonates and Children in Europe: An 11-year Multinational Retrospective Study","volume":"39","author":"Warris","year":"2020","journal-title":"Pediatr. Infect. Dis. J."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1791","DOI":"10.1093\/cid\/ciy776","article-title":"Estimation of Direct Healthcare Costs of Fungal Diseases in the United States","volume":"68","author":"Benedict","year":"2019","journal-title":"Clin. Infect. Dis."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Arastehfar, A., Gabaldon, T., Garcia-Rubio, R., Jenks, J.D., Hoenigl, M., Salzer, H.J.F., Ilkit, M., Lass-Florl, C., and Perlin, D.S. (2020). Drug-Resistant Fungi: An Emerging Challenge Threatening Our Limited Antifungal Armamentarium. Antibiotics, 9.","DOI":"10.3390\/antibiotics9120877"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"165rv113","DOI":"10.1126\/scitranslmed.3004404","article-title":"Hidden killers: Human fungal infections","volume":"4","author":"Brown","year":"2012","journal-title":"Sci Transl. Med."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Basso, V., Tran, D.Q., Ouellette, A.J., and Selsted, M.E. (2020). Host Defense Peptides as Templates for Antifungal Drug Development. J. Fungi, 6.","DOI":"10.3390\/jof6040241"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"105","DOI":"10.3389\/fcimb.2020.00105","article-title":"Antifungal Peptides as Therapeutic Agents","volume":"10","author":"Arbulu","year":"2020","journal-title":"Front. Cell Infect. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1126\/science.abd8700","article-title":"Evolution of fold switching in a metamorphic protein","volume":"371","author":"Dishman","year":"2021","journal-title":"Science"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"5968","DOI":"10.1073\/pnas.1800168115","article-title":"Extant fold-switching proteins are widespread","volume":"115","author":"Porter","year":"2018","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1016\/j.str.2020.10.006","article-title":"Functional and Regulatory Roles of Fold-Switching Proteins","volume":"29","author":"Kim","year":"2021","journal-title":"Structure"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1438","DOI":"10.1021\/acschembio.8b00276","article-title":"Unfolding the Mysteries of Protein Metamorphosis","volume":"13","author":"Dishman","year":"2018","journal-title":"ACS Chem. Biol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2775","DOI":"10.3389\/fimmu.2018.02775","article-title":"A Highly Active Form of XCL1\/Lymphotactin Functions as an Effective Adjuvant to Recruit Cross-Presenting Dendritic Cells for Induction of Effector and Memory CD8(+) T Cells","volume":"9","author":"Matsuo","year":"2018","journal-title":"Front. Immunol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"448","DOI":"10.1189\/jlb.0103024","article-title":"Many chemokines including CCL20\/MIP-3alpha display antimicrobial activity","volume":"74","author":"Yang","year":"2003","journal-title":"J. Leukoc Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"276","DOI":"10.3389\/fimmu.2012.00276","article-title":"Antimicrobial chemokines","volume":"3","author":"Yung","year":"2012","journal-title":"Front. Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"5057","DOI":"10.1073\/pnas.0709518105","article-title":"Interconversion between two unrelated protein folds in the lymphotactin native state","volume":"105","author":"Tuinstra","year":"2008","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2564","DOI":"10.1021\/bi602365d","article-title":"An engineered second disulfide bond restricts lymphotactin\/XCL1 to a chemokine-like conformation with XCR1 agonist activity","volume":"46","author":"Tuinstra","year":"2007","journal-title":"Biochemistry"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2580","DOI":"10.1021\/acschembio.5b00542","article-title":"Engineering Metamorphic Chemokine Lymphotactin\/XCL1 into the GAG-Binding, HIV-Inhibitory Dimer Conformation","volume":"10","author":"Fox","year":"2015","journal-title":"ACS Chem. Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1204","DOI":"10.1021\/acsinfecdis.0c00011","article-title":"Switchable Membrane Remodeling and Antifungal Defense by Metamorphic Chemokine XCL1","volume":"6","author":"Dishman","year":"2020","journal-title":"ACS Infect. Dis."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"3784","DOI":"10.1021\/acs.biochem.6b00353","article-title":"A Requirement for Metamorphic Interconversion in the Antimicrobial Activity of Chemokine XCL1","volume":"55","author":"Nevins","year":"2016","journal-title":"Biochemistry"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"3266","DOI":"10.1016\/j.jmb.2018.06.001","article-title":"The Solution Structure of CCL28 Reveals Structural Lability that Does Not Constrain Antifungal Activity","volume":"430","author":"Thomas","year":"2018","journal-title":"J. Mol. Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1452","DOI":"10.4049\/jimmunol.170.3.1452","article-title":"CCL28 has dual roles in mucosal immunity as a chemokine with broad-spectrum antimicrobial activity","volume":"170","author":"Hieshima","year":"2003","journal-title":"J. Immunol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.mib.2020.03.005","article-title":"Antifungal drug screening: Thinking outside the box to identify novel antifungal scaffolds","volume":"57","author":"Beattie","year":"2020","journal-title":"Curr. Opin. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"657","DOI":"10.1177\/1087057108320713","article-title":"A high-throughput screening assay for small molecules that disrupt yeast cell integrity","volume":"13","author":"Krysan","year":"2008","journal-title":"J. Biomol. Screen"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"He, J., Thomas, M.A., de Anda, J., Lee, M.W., Van Why, E., Simpson, P., Wong, G.C.L., Grayson, M.H., Volkman, B.F., and Huppler, A.R. (2020). Chemokine CCL28 Is a Potent Therapeutic Agent for Oropharyngeal Candidiasis. Antimicrob. Agents Chemother., 64.","DOI":"10.1128\/AAC.00210-20"},{"key":"ref_25","first-page":"3919","article-title":"The antimicrobial peptides and their potential clinical applications","volume":"11","author":"Lei","year":"2019","journal-title":"Am. J. Transl. Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"663","DOI":"10.1080\/14712598.2017.1315402","article-title":"An overview of antimicrobial peptides and the latest advances in their development","volume":"17","author":"Sierra","year":"2017","journal-title":"Expert Opin. Biol. Ther."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1080\/08820139.2019.1655573","article-title":"A Comprehensive Profile of Chemokine Gene Expression in the Tissues of the Female Reproductive Tract in Mice","volume":"49","author":"Menzies","year":"2020","journal-title":"Immunol. Investig."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1007\/s12551-011-0053-8","article-title":"Structural gymnastics of multifunctional metamorphic proteins","volume":"3","author":"Goodchild","year":"2011","journal-title":"Biophys. Rev."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1016\/j.cyto.2014.11.010","article-title":"Structural and agonist properties of XCL2, the other member of the C-chemokine subfamily","volume":"71","author":"Fox","year":"2015","journal-title":"Cytokine"}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/10\/6\/762\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:17:20Z","timestamp":1760163440000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/10\/6\/762"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,6,17]]},"references-count":29,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2021,6]]}},"alternative-id":["pathogens10060762"],"URL":"https:\/\/doi.org\/10.3390\/pathogens10060762","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,6,17]]}}}