{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,10]],"date-time":"2026-08-10T05:30:44Z","timestamp":1786339844026,"version":"build-2736575974"},"reference-count":16,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2026,6,12]],"date-time":"2026-06-12T00:00:00Z","timestamp":1781222400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2026,6,12]],"date-time":"2026-06-12T00:00:00Z","timestamp":1781222400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100009708","name":"Novo Nordisk Fonden","doi-asserted-by":"publisher","award":["NNF21OC0070790"],"award-info":[{"award-number":["NNF21OC0070790"]}],"id":[{"id":"10.13039\/501100009708","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2026,7]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Hydrogenotrophic methanogens are anaerobic archaea that convert carbon dioxide and hydrogen into methane. Central to this process is the heterodisulfide reductase (Hdr), which catalyzes the reduction of the heterodisulfide made of coenzyme M and coenzyme B. In vivo, Hdr functions in association with electron\u2010donating modules such as the [NiFe]\u2010hydrogenase (HdrMvh), which supplies reducing equivalents derived from hydrogen oxidation. Here, we isolate the catalytic properties of Hdr by evaluating substrate turnover independently of its native electron\u2010donating modules using an artificial electron donor system. The molecular features governing substrate recognition and turnover by Hdr were investigated through the design and synthesis of 18 non\u2010endogenous analogs of the native heterodisulfide substrate (compounds\n                    <jats:bold>2a\u2013r<\/jats:bold>\n                    ). We show that several non\u2010endogenous disulfides can serve as alternative substrates for Hdr, including analogs with variation in the coenzyme B\u2010derived amino acid moiety (\n                    <jats:italic>O<\/jats:italic>\n                    \u2010phosphono\u2010Ser,\n                    <jats:italic>L<\/jats:italic>\n                    \u2010Asp,\n                    <jats:italic>L<\/jats:italic>\n                    \u2010Glu, and 2\u2010aminoadipic acid), and that the coenzyme M sulfonate can be replaced by a carboxylate group. In contrast, modifications that disrupt charge balance, chain length, or stereochemical compatibility result in loss of substrate activity and, in some cases, lead to inhibitory behavior, underscoring the narrow constraints required for productive catalysis.\n                  <\/jats:p>","DOI":"10.1002\/pro.70672","type":"journal-article","created":{"date-parts":[[2026,6,12]],"date-time":"2026-06-12T11:42:26Z","timestamp":1781264546000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Structure\u2013activity relationship study shows versatility in substrate specificity for heterodisulfide reductase in methanogenic archaea"],"prefix":"10.1002","volume":"35","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5283-9196","authenticated-orcid":false,"given":"Mahdi Faal","family":"Maleki","sequence":"first","affiliation":[{"name":"Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences University of Copenhagen  Frederiksberg Denmark"},{"name":"Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences University of Copenhagen  Copenhagen Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4369-3858","authenticated-orcid":false,"given":"Joakim B\u00f8gelund","family":"Jakobsen","sequence":"additional","affiliation":[{"name":"Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences University of Copenhagen  Copenhagen Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva Danielle","family":"Schuiten","sequence":"additional","affiliation":[{"name":"Max Planck Institute for Marine Microbiology  Bremen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tristan","family":"Wagner","sequence":"additional","affiliation":[{"name":"Max Planck Institute for Marine Microbiology  Bremen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andr\u00e9 Luis Alves","family":"Neves","sequence":"additional","affiliation":[{"name":"Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences University of Copenhagen  Frederiksberg Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lennart","family":"Bunch","sequence":"additional","affiliation":[{"name":"Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences University of Copenhagen  Copenhagen Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,6,12]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.accounts.4c00730"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi700679u"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(89)81062-2"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1990.tb19331.x"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41589-022-01232-y"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41564-023-01398-8"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1016761108"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-020-10724-7"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41589-025-02055-3"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.biochem.2c00135"},{"key":"e_1_2_10_12_1","first-page":"75","volume-title":"Biochemical preparations","author":"Neuhaus FC","year":"1958"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-025-09229-y"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.biochem.9b00164"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro1931"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.aan0425"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.abg5550"}],"updated-by":[{"DOI":"10.1002\/pro.70711","type":"correction","label":"Correction","source":"publisher","updated":{"date-parts":[[2026,7,12]],"date-time":"2026-07-12T00:00:00Z","timestamp":1783814400000}}],"container-title":["Protein Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pro.70672","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/pro.70672","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pro.70672","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,10]],"date-time":"2026-08-10T05:25:37Z","timestamp":1786339537000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/pro.70672"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,12]]},"references-count":16,"journal-issue":{"issue":"7","published-print":{"date-parts":[[2026,7]]}},"alternative-id":["10.1002\/pro.70672"],"URL":"https:\/\/doi.org\/10.1002\/pro.70672","archive":["Portico"],"relation":{},"ISSN":["0961-8368","1469-896X"],"issn-type":[{"value":"0961-8368","type":"print"},{"value":"1469-896X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,12]]},"assertion":[{"value":"2025-12-29","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication 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