{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,28]],"date-time":"2026-03-28T04:28:16Z","timestamp":1774672096671,"version":"3.50.1"},"reference-count":34,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2022,5,26]],"date-time":"2022-05-26T00:00:00Z","timestamp":1653523200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002661","name":"Research Foundation for Industry and Agriculture (FRIA)","doi-asserted-by":"publisher","award":["FRIA1.E049.16"],"award-info":[{"award-number":["FRIA1.E049.16"]}],"id":[{"id":"10.13039\/501100002661","id-type":"DOI","asserted-by":"publisher"}]},{"name":"FEDER and Wallonia (BIOMED HUB Technology Support project)","award":["FRIA1.E049.16"],"award-info":[{"award-number":["FRIA1.E049.16"]}]},{"name":"Belgian Fund for Scientific Research (F.R.S.-FNRS)","award":["FRIA1.E049.16"],"award-info":[{"award-number":["FRIA1.E049.16"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Ferroverdins are ferrous iron (Fe2+)-nitrosophenolato complexes produced by a few Streptomyces species as a response to iron overload. Previously, three ferroverdins were identified: ferroverdin A, in which three molecules of p-vinylphenyl-3-nitroso-4-hydroxybenzoate (p-vinylphenyl-3,4-NHBA) are recruited to bind Fe2+, and Ferroverdin B and Ferroverdin C, in which one molecule of p-vinylphenyl-3,4-NHBA is substituted by hydroxy-p-vinylphenyl-3,4-NHBA, and by carboxy-p-vinylphenyl-3,4-NHBA, respectively. These molecules, especially ferroverdin B, are potent inhibitors of the human cholesteryl ester transfer protein (CETP) and therefore candidate hits for the development of drugs that increase the serum concentration of high-density lipoprotein cholesterol, thereby diminishing the risk of atherosclerotic cardiovascular disease. In this work, we used high-resolution mass spectrometry combined with tandem mass spectrometry to identify 43 novel ferroverdins from the cytosol of two Streptomyces lunaelactis species. For 13 of them (designated ferroverdins C2, C3, D, D2, D3, E, F, G, H, CD, DE, DF, and DG), we could elucidate their structure, and for the other 17 new ferroverdins, ambiguity remains for one of the three ligands. p-formylphenyl-3,4-NHBA, p-benzoic acid-3,4-NHBA, 3,4-NHBA, p-phenylpropionate-3,4-NHBA, and p-phenyacetate-3,4-NHBA were identified as new alternative chelators for Fe2+-binding, and two compounds (C3 and D3) are the first reported ferroverdins that do not recruit p-vinylphenyl-3,4-NHBA. Our work thus uncovered putative novel CETP inhibitors or ferroverdins with novel bioactivities.<\/jats:p>","DOI":"10.3390\/biom12060752","type":"journal-article","created":{"date-parts":[[2022,5,26]],"date-time":"2022-05-26T23:04:31Z","timestamp":1653606271000},"page":"752","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Structure of New Ferroverdins Recruiting Unconventional Ferrous Iron Chelating Agents"],"prefix":"10.3390","volume":"12","author":[{"given":"Lo\u00efc","family":"Martinet","sequence":"first","affiliation":[{"name":"InBioS, Center for Protein Engineering, University of Liege, B-4000 Liege, Belgium"},{"name":"Hedera-22, Boulevard du Rectorat 27b, B-4000 Liege, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1622-0118","authenticated-orcid":false,"given":"Dominique","family":"Baiwir","sequence":"additional","affiliation":[{"name":"GIGA Proteomics Facility, University of Liege, B-4000 Liege, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8757-8133","authenticated-orcid":false,"given":"Gabriel","family":"Mazzucchelli","sequence":"additional","affiliation":[{"name":"Mass Spectrometry Laboratory, MolSys Research Unit, University of Liege, B-4000 Liege, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4022-7325","authenticated-orcid":false,"given":"S\u00e9bastien","family":"Rigali","sequence":"additional","affiliation":[{"name":"InBioS, Center for Protein Engineering, University of Liege, B-4000 Liege, Belgium"},{"name":"Hedera-22, Boulevard du Rectorat 27b, B-4000 Liege, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,5,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Nicholls, A.J., Barber, T., and Baxendale, I.R. (2019). The Synthesis and Utility of Metal-Nitrosophenolato Compounds\u2014Highlighting the Baudisch Reaction. Molecules, 24.","DOI":"10.3390\/molecules24224018"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"645","DOI":"10.1038\/176645a0","article-title":"Ferroverdin, a Green Pigment Containing Iron Produced by a Streptomycete","volume":"176","author":"Chain","year":"1955","journal-title":"Nature"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1007\/s10482-014-0348-4","article-title":"Streptomyces lunaelactis sp. Nov., a Novel Ferroverdin A-Producing Streptomyces Species Isolated from a Moonmilk Speleothem","volume":"107","author":"Maciejewska","year":"2015","journal-title":"Antonie Van Leeuwenhoek"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Martinet, L., Na\u00f4m\u00e9, A., Baiwir, D., De Pauw, E., Mazzucchelli, G., and Rigali, S. (2020). On the Risks of Phylogeny-Based Strain Prioritization for Drug Discovery: Streptomyces lunaelactis as a Case Study. Biomolecules, 10.","DOI":"10.3390\/biom10071027"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Adam, D., Maciejewska, M., Na\u00f4m\u00e9, A., Martinet, L., Coppieters, W., Karim, L., Baurain, D., and Rigali, S. (2018). Isolation, Characterization, and Antibacterial Activity of Hard-to-Culture Actinobacteria from Cave Moonmilk Deposits. Antibiotics, 7.","DOI":"10.3390\/antibiotics7020028"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1455","DOI":"10.3389\/fmicb.2016.01455","article-title":"A Phenotypic and Genotypic Analysis of the Antimicrobial Potential of Cultivable Streptomyces Isolated from Cave Moonmilk Deposits","volume":"7","author":"Maciejewska","year":"2016","journal-title":"Front. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Maciejewska, M., Ca\u0142usi\u0144ska, M., Cornet, L., Adam, D., Pessi, I.S., Malchair, S., Delfosse, P., Baurain, D., Barton, H.A., and Carnol, M. (2018). High-Throughput Sequencing Analysis of the Actinobacterial Spatial Diversity in Moonmilk Deposits. Antibiotics, 7.","DOI":"10.20944\/preprints201802.0090.v1"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"589","DOI":"10.1038\/224589a0","article-title":"Structure of Ferroverdin","volume":"224","author":"Candeloro","year":"1969","journal-title":"Nature"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"769","DOI":"10.1038\/194769b0","article-title":"Structure of Ferroverdin","volume":"194","author":"Ballio","year":"1962","journal-title":"Nature"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1101","DOI":"10.7164\/antibiotics.52.1101","article-title":"Ferroverdins, Inhibitors of Cholesteryl Ester Transfer Protein Produced by Streptomyces sp. WK-5344. I. Production, Isolation and Biological Properties","volume":"52","author":"Tomoda","year":"1999","journal-title":"J. Antibiot."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1128\/jb.149.1.381-383.1982","article-title":"Production of Deferriferrioxamines B and E from a Ferroverdin-Producing Streptomyces Species","volume":"149","author":"Yang","year":"1982","journal-title":"J. Bacteriol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e01230-19","DOI":"10.1128\/mBio.01230-19","article-title":"A Single Biosynthetic Gene Cluster Is Responsible for the Production of Bagremycin Antibiotics and Ferroverdin Iron Chelators","volume":"10","author":"Martinet","year":"2019","journal-title":"mBio"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"628","DOI":"10.1111\/mmi.12008","article-title":"Interspecies Modulation of Bacterial Development through Iron Competition and Siderophore Piracy","volume":"86","author":"Traxler","year":"2012","journal-title":"Mol. Microbiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1390","DOI":"10.1039\/C4MT00068D","article-title":"Altered Desferrioxamine-Mediated Iron Utilization Is a Common Trait of Bald Mutants of Streptomyces coelicolor","volume":"6","author":"Lambert","year":"2014","journal-title":"Metallomics"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2899","DOI":"10.1099\/mic.0.28139-0","article-title":"Desferrioxamine E Produced by Streptomyces Griseus Stimulates Growth and Development of Streptomyces tanashiensis","volume":"151","author":"Yamanaka","year":"2005","journal-title":"Microbiology"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"512","DOI":"10.1111\/j.1758-2229.2012.00354.x","article-title":"Unsuspected Control of Siderophore Production by N-Acetylglucosamine in Streptomycetes","volume":"4","author":"Craig","year":"2012","journal-title":"Environ. Microbiol. Rep."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1111\/j.1751-7915.2010.00240.x","article-title":"Disruption of the Siderophore-Binding DesE Receptor Gene in Streptomyces coelicolor A3(2) Results in Impaired Growth in Spite of Multiple Iron-Siderophore Transport Systems","volume":"4","author":"Gosset","year":"2011","journal-title":"Microb. Biotechnol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"e00459-13","DOI":"10.1128\/mBio.00459-13","article-title":"Interspecies Interactions Stimulate Diversification of the Streptomyces coelicolor Secreted Metabolome","volume":"4","author":"Traxler","year":"2013","journal-title":"mBio"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e01503-18","DOI":"10.1128\/AEM.01503-18","article-title":"IdeR, a DtxR Family Iron Response Regulator, Controls Iron Homeostasis, Morphological Differentiation, Secondary Metabolism, and the Oxidative Stress Response in Streptomyces avermitilis","volume":"84","author":"Cheng","year":"2018","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1742","DOI":"10.3389\/fmicb.2018.01742","article-title":"Production of Prodiginines Is Part of a Programmed Cell Death Process in Streptomyces coelicolor","volume":"9","author":"Tenconi","year":"2018","journal-title":"Front. Microbiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e00435-18","DOI":"10.1128\/genomeA.00435-18","article-title":"Complete Genome Sequence of Streptomyces lunaelactis MM109T, Isolated from Cave Moonmilk Deposits","volume":"6","author":"Maciejewska","year":"2018","journal-title":"Genome Announc."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"482","DOI":"10.1080\/09168451.2018.1553605","article-title":"Characterization of the Bagremycin Biosynthetic Gene Cluster in Streptomyces sp. T\u00fc 4128","volume":"83","author":"Ye","year":"2019","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1108","DOI":"10.7164\/antibiotics.52.1108","article-title":"Ferroverdins, Inhibitors of Cholesteryl Ester Transfer Protein Produced by Streptomyces sp. WK-5344. II. Structure Elucidation","volume":"52","author":"Tabata","year":"1999","journal-title":"J. Antibiot."},{"key":"ref_24","unstructured":"Omura, S., Tomoda, H., and Takahashi, Y. (2003). Substances Wk-5344a and Wk-5344b and Process for Producing the Same. (6,512,008), U.S. Patent."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1080\/14779072.2020.1745633","article-title":"The Mystery of Evacetrapib-Why Are CETP Inhibitors Failing?","volume":"18","author":"Nicholls","year":"2020","journal-title":"Expert Rev. Cardiovasc. Ther."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"947","DOI":"10.1001\/jama.2017.11467","article-title":"Association of Genetic Variants Related to CETP Inhibitors and Statins With Lipoprotein Levels and Cardiovascular Risk","volume":"318","author":"Ference","year":"2017","journal-title":"JAMA"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"37","DOI":"10.4137\/CMC.S32667","article-title":"CETP Inhibition: Past Failures and Future Hopes","volume":"10","author":"Kosmas","year":"2016","journal-title":"Clin. Med. Insights Cardiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1161\/CIRCRESAHA.117.311978","article-title":"The Trials and Tribulations of CETP Inhibitors","volume":"122","author":"Tall","year":"2018","journal-title":"Circ. Res."},{"key":"ref_29","unstructured":"Kieser, T., Bibb, M.J., Buttner, M.J., Chater, K.F., and Hopwood, D.A. (2000). Practical Streptomyces Genetics, John Innes Foundation."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"11298","DOI":"10.1021\/ac503000e","article-title":"ChelomEx: Isotope-Assisted Discovery of Metal Chelates in Complex Media Using High-Resolution LC-MS","volume":"86","author":"Baars","year":"2014","journal-title":"Anal. Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1128\/AEM.03160-15","article-title":"The Siderophore Metabolome of Azotobacter vinelandii","volume":"82","author":"Baars","year":"2016","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"6096","DOI":"10.1039\/C4AN01461H","article-title":"Metallophore Mapping in Complex Matrices by Metal Isotope Coded Profiling of Organic Ligands","volume":"139","author":"Deicke","year":"2014","journal-title":"Analyst"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1128\/AEM.02339-12","article-title":"Isotope-Assisted Screening for Iron-Containing Metabolites Reveals a High Degree of Diversity among Known and Unknown Siderophores Produced by Trichoderma spp.","volume":"79","author":"Lehner","year":"2013","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Hamdali, H., Lebrihi, A., Monje, M.C., Benharref, A., Hafidi, M., Ouhdouch, Y., and Virolle, M.J. (2021). A Molecule of the Viridomycin Family Originating from a Streptomyces griseus-Related Strain Has the Ability to Solubilize Rock Phosphate and to Inhibit Microbial Growth. Antibiotics, 10.","DOI":"10.3390\/antibiotics10010072"}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/12\/6\/752\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:19:24Z","timestamp":1760138364000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/12\/6\/752"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,5,26]]},"references-count":34,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2022,6]]}},"alternative-id":["biom12060752"],"URL":"https:\/\/doi.org\/10.3390\/biom12060752","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,5,26]]}}}