{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,5]],"date-time":"2026-06-05T01:53:14Z","timestamp":1780624394183,"version":"3.54.1"},"reference-count":39,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2021,12,18]],"date-time":"2021-12-18T00:00:00Z","timestamp":1639785600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100010713","name":"Indian Institute of Science Education and Research Bhopal","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100010713","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2022,3]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Accumulation of formaldehyde, a highly reactive molecule, in the cell is toxic, and requires detoxification for the organism's survival. Mycothiol\u2010dependent formaldehyde dehydrogenase or <jats:italic>S<\/jats:italic>\u2010nitrosomycothiol reductase (MscR) from <jats:italic>Mycobacterium smegmatis<\/jats:italic> and <jats:italic>Mycobacterium tuberculosis<\/jats:italic> was previously known for detoxifying formaldehyde and protecting the cell against nitrosative stress. We here show that <jats:italic>M. smegmatis<\/jats:italic> MscR exhibits a mycothiol\u2010independent formaldehyde dehydrogenase (FDH) activity <jats:italic>in vitro<\/jats:italic>. Presence of zinc in the reaction enhances MscR activity, thus making it a zinc\u2010dependent FDH. Interestingly, MscR utilizes only formaldehyde and no other primary aldehydes as its substrate <jats:italic>in vitro<\/jats:italic>, and <jats:italic>M. smegmatis<\/jats:italic> lacking <jats:italic>mscR<\/jats:italic> (\u0394<jats:italic>mscR<\/jats:italic>) shows sensitivity exclusively toward formaldehyde. Bioinformatics analysis of MscRs from various bacteria reveals 10 positionally conserved cysteines, whose importance in structural stability and biological activity is not yet investigated. To explore the significance of these cysteines, we generated MscR single Cys variants by systematically replacing each cysteine with serine. All of the Cys variants except C39S and C309S are unable to show a complete rescue of \u0394<jats:italic>mscR<\/jats:italic> on formaldehyde, show a significant loss of enzymatic activity <jats:italic>in vitro<\/jats:italic>, pronounced structural alterations as probed by circular dichroism, and loss of homotetramerization on size exclusion chromatography. Our data thus reveal the importance of intact cysteines in the structural stability and biological activity of MscR, which is a dedicated FDH in <jats:italic>M. smegmatis<\/jats:italic>, and shows ~84% identity with <jats:italic>M. tuberculosis<\/jats:italic> MscR. We believe that this knowledge will further help in the development of FDH as a potential drug target against <jats:italic>M. tuberculosis<\/jats:italic> infections.<\/jats:p>","DOI":"10.1002\/pro.4258","type":"journal-article","created":{"date-parts":[[2021,12,14]],"date-time":"2021-12-14T07:35:39Z","timestamp":1639467339000},"page":"628-638","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Molecular dissection of a dedicated formaldehyde dehydrogenase from <scp><i>Mycobacterium smegmatis<\/i><\/scp>"],"prefix":"10.1002","volume":"31","author":[{"given":"Saloni Rajesh","family":"Wani","sequence":"first","affiliation":[{"name":"Microbiology and Molecular Biology Laboratory, Department of Biological Sciences Indian Institute of Science Education and Research (IISER)  Bhopal 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