{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T07:06:59Z","timestamp":1790838419644,"version":"4.1.0"},"reference-count":35,"publisher":"Oxford University Press (OUP)","issue":"10","license":[{"start":{"date-parts":[[2022,1,1]],"date-time":"2022-01-01T00:00:00Z","timestamp":1640995200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"DOI":"10.13039\/100009007","name":"Health Research Foundation","doi-asserted-by":"publisher","award":["MOP-142421"],"award-info":[{"award-number":["MOP-142421"]}],"id":[{"id":"10.13039\/100009007","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2022,10,8]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>[NiFe]-hydrogenases are used by several human pathogens to catalyze the reversible conversion between molecular hydrogen and protons and electrons. Hydrogenases provide an increased metabolic flexibility for pathogens, such as Escherichia coli and Helicobacter pylori, by allowing the use of molecular hydrogen as an energy source to promote survival in anaerobic environments. With the rise of antimicrobial resistance and the desire for novel therapeutics, the [NiFe]-hydrogenases are alluring targets. Inhibiting the nickel insertion pathway of [NiFe]-hydrogenases is attractive as this pathway is required for the generation of functional enzymes and is orthogonal to human biochemistry. In this work, nickel availability for the production and function of E. coli [NiFe]-hydrogenase was explored through immunoblot and activity assays. Whole-cell hydrogenase activities were assayed in high throughput against a small molecule library of known bioactives. Iodoquinol was identified as a potential inhibitor of the nickel biosynthetic pathway of [NiFe]-hydrogenase through a two-step screening process, but further studies with immunoblot assays showed confounding effects dependent on the cell growth phase. This study highlights the significance of considering the growth phenotype for whole-cell based assays overall and its effects on various cellular processes influenced by metal trafficking and homeostasis.<\/jats:p>","DOI":"10.1093\/mtomcs\/mfac073","type":"journal-article","created":{"date-parts":[[2022,10,3]],"date-time":"2022-10-03T13:34:19Z","timestamp":1664804059000},"source":"Crossref","is-referenced-by-count":5,"title":["Using a high-throughput, whole-cell hydrogenase assay to identify potential small molecule inhibitors of [NiFe]-hydrogenase"],"prefix":"10.1093","volume":"14","author":[{"given":"Stephanie","family":"Sebastiampillai","sequence":"first","affiliation":[{"name":"Department of Chemistry, University of Toronto , Toronto, Ontario M5S 3H6 , Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael J","family":"Lacasse","sequence":"additional","affiliation":[{"name":"Department of Chemistry, University of Toronto , Toronto, Ontario M5S 3H6 , Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susan","family":"McCusker","sequence":"additional","affiliation":[{"name":"Centre for Microbial Chemical Biology , MDCL-2330, Hamilton, Ontario L8S 4K1 , Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tracey","family":"Campbell","sequence":"additional","affiliation":[{"name":"Centre for Microbial Chemical Biology , MDCL-2330, Hamilton, Ontario L8S 4K1 , Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark","family":"Nitz","sequence":"additional","affiliation":[{"name":"Department of Chemistry, University of Toronto , Toronto, Ontario M5S 3H6 , Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deborah B","family":"Zamble","sequence":"additional","affiliation":[{"name":"Department of Chemistry, University of Toronto , Toronto, Ontario M5S 3H6 , Canada"},{"name":"Department of Biochemistry, University of Toronto , Toronto, Ontario M5S 1A8 , Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2022,10,3]]},"reference":[{"key":"2024052508134098400_bib1","doi-asserted-by":"crossref","first-page":"1689","DOI":"10.1021\/acs.biochem.5b01328","article-title":"[NiFe]-Hydrogenase Maturation","volume":"55","author":"Lacasse","year":"2016","journal-title":"Biochemistry"},{"key":"2024052508134098400_bib2","volume-title":"WHO publishes list of bacteria for which new antibiotics are urgently needed","author":"WHO","year":"2017"},{"key":"2024052508134098400_bib3","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1111\/j.1574-6968.1996.tb08365.x","article-title":"Hydrogen uptake hydrogenase in Helicobacter pylori","volume":"141","author":"Maier","year":"1996","journal-title":"FEMS Microbiol. 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