{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T14:11:31Z","timestamp":1781619091026,"version":"3.54.5"},"reference-count":49,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2020,3,16]],"date-time":"2020-03-16T00:00:00Z","timestamp":1584316800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006188","name":"Einstein Stiftung Berlin","doi-asserted-by":"publisher","award":["EVF\u20102016\u2010277"],"award-info":[{"award-number":["EVF\u20102016\u2010277"]}],"id":[{"id":"10.13039\/501100006188","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["MicrobiologyOpen"],"published-print":{"date-parts":[[2020,6]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>[NiFe]\u2010hydrogenases catalyze the reversible conversion of molecular hydrogen into protons end electrons. This reaction takes place at a NiFe(CN)<jats:sub>2<\/jats:sub>(CO) cofactor located in the large subunit of the bipartite hydrogenase module. The corresponding apo\u2010protein carries usually a C\u2010terminal extension that is cleaved off by a specific endopeptidase as soon as the cofactor insertion has been accomplished by the maturation machinery. This process triggers complex formation with the small, electron\u2010transferring subunit of the hydrogenase module, revealing catalytically active enzyme. The role of the C\u2010terminal extension in cofactor insertion, however, remains elusive. We have addressed this problem by using genetic engineering to remove the entire C\u2010terminal extension from the apo\u2010form of the large subunit of the membrane\u2010bound [NiFe]\u2010hydrogenase (MBH) from <jats:italic>Ralstonia eutropha<\/jats:italic>. Unexpectedly, the MBH holoenzyme derived from this precleaved large subunit was targeted to the cytoplasmic membrane, conferred H<jats:sub>2<\/jats:sub>\u2010dependent growth of the host strain, and the purified protein showed exactly the same catalytic activity as native MBH. The only difference was a reduced hydrogenase content in the cytoplasmic membrane. These results suggest that in the case of the <jats:italic>R.\u00a0eutropha<\/jats:italic> MBH, the C\u2010terminal extension is dispensable for cofactor insertion and seems to function only as a maturation facilitator.<\/jats:p>","DOI":"10.1002\/mbo3.1029","type":"journal-article","created":{"date-parts":[[2020,3,17]],"date-time":"2020-03-17T06:32:35Z","timestamp":1584426755000},"page":"1197-1206","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["A membrane\u2010bound [NiFe]\u2010hydrogenase large subunit precursor whose C\u2010terminal extension is not essential for cofactor incorporation but guarantees optimal maturation"],"prefix":"10.1002","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1395-938X","authenticated-orcid":false,"given":"Sven","family":"Hartmann","sequence":"first","affiliation":[{"name":"Institut f\u00fcr Chemie Physikalische Chemie Technische Universit\u00e4t Berlin Berlin Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4141-7836","authenticated-orcid":false,"given":"Stefan","family":"Frielingsdorf","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Chemie Physikalische Chemie Technische Universit\u00e4t Berlin Berlin Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0986-3059","authenticated-orcid":false,"given":"Giorgio","family":"Caserta","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Chemie Physikalische Chemie Technische Universit\u00e4t Berlin Berlin Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4550-5128","authenticated-orcid":false,"given":"Oliver","family":"Lenz","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Chemie Physikalische Chemie Technische Universit\u00e4t Berlin Berlin 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