A Reduced F420-Dependent Nitrite Reductase in an Anaerobic Methanotrophic Archaeon

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dc.contributor.authorHeryakusuma, Christianen
dc.contributor.authorSusanti, Dwien
dc.contributor.authorYu, Hangen
dc.contributor.authorLi, Zhouen
dc.contributor.authorPurwantini, Endangen
dc.contributor.authorHettich, Robert L.en
dc.contributor.authorOrphan, Victoria J.en
dc.contributor.authorMukhopadhyay, Biswarupen
dc.date.accessioned2023-01-17T20:53:02Zen
dc.date.available2023-01-17T20:53:02Zen
dc.date.issued2022-06-13en
dc.date.updated2023-01-17T16:12:01Zen
dc.description.abstractAnaerobic methanotrophic archaea (ANME), which oxidize methane in marine sediments through syntrophic associations with sulfate-reducing bacteria, carry homologs of coenzyme F420-dependent sulfite reductase (Fsr) of Methanocaldococcus jannaschii, a hyperthermophilic methanogen from deep-sea hydrothermal vents. M. jannaschii Fsr (MjFsr) and ANME-Fsr belong to two phylogenetically distinct groups, FsrI and FsrII, respectively. MjFsrI reduces sulfite to sulfide with reduced F420 (F420H2), protecting methyl coenzyme M reductase (Mcr), an essential enzyme for methanogens, from sulfite inhibition. However, the function of FsrIIs in ANME, which also rely on Mcr and live in sulfidic environments, is unknown. We have determined the catalytic properties of FsrII from a member of ANME-2c. Since ANME remain to be isolated, we expressed ANME2c-FsrII in a closely related methanogen, Methanosarcina acetivorans. Purified recombinant FsrII contained siroheme, indicating that the methanogen, which lacks a native sulfite reductase, produced this coenzyme. Unexpectedly, FsrII could not reduce sulfite or thiosulfate with F420H2. Instead, it acted as an F420H2-dependent nitrite reductase (FNiR) with physiologically relevant Km values (nitrite, 5 μM; F420H2, 14 μM). From kinetic, thermodynamic, and structural analyses, we hypothesize that in FNiR, F420H2- derived electrons are delivered at the oxyanion reduction site at a redox potential that is suitable for reducing nitrite (E09 [standard potential], 1440 mV) but not sulfite (E09, 2116 mV). These findings and the known nitrite sensitivity of Mcr suggest that FNiR may protect nondenitrifying ANME from nitrite toxicity. Remarkably, by reorganizing the reductant processing system, Fsr transforms two analogous oxyanions in two distinct archaeal lineages with different physiologies and ecologies. IMPORTANCE Coenzyme F420-dependent sulfite reductase (Fsr) protects methanogenic archaea inhabiting deep-sea hydrothermal vents from the inactivation of methyl coenzyme M reductase (Mcr), one of their essential energy production enzymes. Anaerobic methanotrophic archaea (ANME) that oxidize methane and rely on Mcr, carry Fsr homologs that form a distinct clade. We show that a member of this clade from ANME-2c functions as F420-dependent nitrite reductase (FNiR) and lacks Fsr activity. This specialization arose from a distinct feature of the reductant processing system and not the substrate recognition element. We hypothesize FNiR may protect ANME Mcr from inactivation by nitrite. This is an example of functional specialization within a protein family that is induced by changes in electron transfer modules to fit an ecological need.en
dc.description.versionPublished versionen
dc.format.extent21 page(s)en
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.1128/jb.00078-22en
dc.identifier.eissn1098-5530en
dc.identifier.issn0021-9193en
dc.identifier.issue7en
dc.identifier.orcidMukhopadhyay, Biswarup [0000-0003-0736-0298]en
dc.identifier.orcidPurwantini, Endang [0000-0002-4113-9995]en
dc.identifier.pmid35695516en
dc.identifier.urihttp://hdl.handle.net/10919/113207en
dc.identifier.volume204en
dc.language.isoenen
dc.publisherAmerican Society for Microbiologyen
dc.relation.urihttp://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000811638400001&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=930d57c9ac61a043676db62af60056c1en
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectanaerobic methane oxidationen
dc.subjectanaerobic methanotrophic archaeaen
dc.subjectmethanogenen
dc.subjectF-420-dependent nitrite reductaseen
dc.subjectF-420-dependent sulfite reductaseen
dc.subjectFsrIen
dc.subjectFsrIIen
dc.subjectcoenzyme F-420en
dc.subjectF420H2en
dc.subjectdeazaflavinen
dc.subjectelectron transferen
dc.subjectiron-sulfur clusteren
dc.subjectmethaneen
dc.subjectDISSIMILATORY SULFITE REDUCTASEen
dc.subjectMULTIPLE SEQUENCE ALIGNMENTen
dc.subjectCOENZYME-M REDUCTASEen
dc.subjectMETHANE OXIDATIONen
dc.subjectBIOCHEMICAL-CHARACTERIZATIONen
dc.subjectMETHANOSARCINA-BARKERIen
dc.subjectF420H2 DEHYDROGENASEen
dc.subjectELECTRON-TRANSFERen
dc.subjectPURIFICATIONen
dc.subjectSULFURen
dc.subjectF420-dependent nitrite reductaseen
dc.subjectF420-dependent sulfite reductaseen
dc.subjectcoenzyme F420en
dc.subject14 Life Below Wateren
dc.subject.meshArchaeaen
dc.subject.meshNitritesen
dc.subject.meshMethaneen
dc.subject.meshNitrite Reductasesen
dc.subject.meshRiboflavinen
dc.subject.meshReducing Agentsen
dc.subject.meshAnaerobiosisen
dc.subject.meshOxidation-Reductionen
dc.subject.meshOxidoreductases Acting on Sulfur Group Donorsen
dc.titleA Reduced F<sub>420</sub>-Dependent Nitrite Reductase in an Anaerobic Methanotrophic Archaeonen
dc.title.serialJournal of Bacteriologyen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten
dc.type.otherArticleen
dc.type.otherEarly Accessen
dc.type.otherJournalen
pubs.organisational-group/Virginia Techen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciencesen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciences/Biochemistryen
pubs.organisational-group/Virginia Tech/Faculty of Health Sciencesen
pubs.organisational-group/Virginia Tech/All T&R Facultyen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciences/CALS T&R Facultyen
pubs.organisational-group/Virginia Tech/VT Carilion School of Medicineen
pubs.organisational-group/Virginia Tech/VT Carilion School of Medicine/Internal Medicineen
pubs.organisational-group/Virginia Tech/VT Carilion School of Medicine/Internal Medicine/Internal Med-Subgroupen

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