{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T03:27:10Z","timestamp":1783567630504,"version":"3.55.0"},"reference-count":55,"publisher":"Portland Press Ltd.","issue":"2","content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2014,12,1]]},"abstract":"<jats:p>[NiFe]-hydrogenases (Hyd) bind a nickel-iron-based cofactor. The Fe ion of the cofactor is bound by two cyanide ligands and a single carbon monoxide ligand. Minimally six accessory proteins (HypA\u2013HypF) are necessary for NiFe(CN)2CO cofactor biosynthesis in Escherichia coli. It has been shown that the anaerobically purified HypC\u2013HypD\u2013HypE scaffold complex carries the Fe(CN)2CO moiety of this cofactor. In the present study, we have purified the HybG\u2013HypDE complex and used it to successfully reconstitute in\u00a0vitro active Hyd from E. coli. HybG is a homologue of HypC that is specifically required for the maturation of Hyd-2 and also functions in the maturation of Hyd-1 of E. coli. Maturation of active Hyd-1 and Hyd-2 could be demonstrated in extracts derived from HybG- and HypD-deficient E. coli strains by adding anaerobically purified HybG\u2013HypDE complex. In\u00a0vitro maturation was dependent on ATP, carbamoylphosphate, nickel and reducing conditions. Hydrogenase maturation was prevented when the purified HybG\u2013HypDE complex used in the maturation assay lacked a bound Fe(CN)2CO moiety. These findings demonstrate that it is possible to isolate incompletely processed intermediates on the maturation pathway and to use these to activate apo-forms of [NiFe]-hydrogenase large subunits.<\/jats:p>","DOI":"10.1042\/bj20140485","type":"journal-article","created":{"date-parts":[[2014,9,4]],"date-time":"2014-09-04T17:16:39Z","timestamp":1409850999000},"page":"169-177","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":22,"title":["[NiFe]-hydrogenase maturation <i>in\u00a0vitro<\/i>: analysis of the roles of the HybG and HypD accessory proteins"],"prefix":"10.1042","volume":"464","author":[{"given":"Basem","family":"Soboh","sequence":"first","affiliation":[{"name":"Institute of Microbiology, Martin-Luther University Halle-Wittenberg, Kurt-Mothes-Str. 3, 06120 Halle (Saale), Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ute","family":"Lindenstrauss","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, Martin-Luther University Halle-Wittenberg, Kurt-Mothes-Str. 3, 06120 Halle (Saale), Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudia","family":"Granich","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, Martin-Luther University Halle-Wittenberg, Kurt-Mothes-Str. 3, 06120 Halle (Saale), Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mahwish","family":"Javed","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, Martin-Luther University Halle-Wittenberg, Kurt-Mothes-Str. 3, 06120 Halle (Saale), Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin","family":"Herzberg","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, Martin-Luther University Halle-Wittenberg, Kurt-Mothes-Str. 3, 06120 Halle (Saale), Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudia","family":"Thomas","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, Martin-Luther University Halle-Wittenberg, Kurt-Mothes-Str. 3, 06120 Halle (Saale), Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sven\u00a0T.","family":"Stripp","sequence":"additional","affiliation":[{"name":"Freie Universit\u00e4t Berlin, Experimental Molecular Biophysics, Arnimalle 14, 14195 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"288","published-online":{"date-parts":[[2014,11,14]]},"reference":[{"key":"2021112216113519000_B1","doi-asserted-by":"publisher","first-page":"4206","DOI":"10.1021\/cr050196r","article-title":"Occurrence, classification, and biological function of hydrogenases: an overview","volume":"107","author":"Vignais","year":"2007","journal-title":"Chem. 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