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A two-component protease in Methylorubrum extorquens with high activity toward the peptide precursor of the redox cofactor pyrroloquinoline quinone

2019-10Journal article Open access
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dc.contributor.authorMartins, Ana M.
dc.contributor.authorLatham, John A.
dc.contributor.authorMartel, Paulo
dc.contributor.authorBarr, Ian
dc.contributor.authorIavarone, Anthony T.
dc.contributor.authorKlinman, Judith P.
dc.date.accessioned2020-07-24T10:51:20Z
dc.date.available2020-07-24T10:51:20Z
dc.date.issued2019-10
dc.description.abstractPyrroloquinoline quinone is a prominent redox cofactor in many prokaryotes, produced from a ribosomally synthesized and post-translationally modified peptide PqqA via a pathway comprising four conserved proteins PqqB?E. These four proteins are now fairly well-characterized and span radical SAM activity (PqqE), aided by a peptide chaperone (PqqD), a dual hydroxylase (PqqB), and an eight-electron, eight-proton oxidase (PqqC). A full description of this pathway has been hampered by a lack of information regarding a protease/peptidase required for the excision of an early, cross-linked di-amino acid precursor to pyrroloquinoline quinone. Herein, we isolated and characterized a two-component heterodimer protein from the ?-proteobacterium Methylobacterium (Methylorubrum) extorquens that can rapidly catalyze cleavage of PqqA into smaller peptides. Using pulldown assays, surface plasmon resonance, and isothermal calorimetry, we demonstrated the formation of a complex PqqF/PqqG, with a K-D of 300 nm. We created a molecular model of the heterodimer by comparison with the Sphingomonas sp. A1 M16B Sph2681/Sph2682 protease. Analysis of time-dependent patterns for the appearance of proteolysis products indicates high specificity of PqqF/PqqG for serine side chains. We hypothesize that PqqF/PqqG initially cleaves between the PqqE/PqqD-generated cross-linked form of PqqA, with nonspecific cellular proteases completing the release of a suitable substrate for the downstream enzyme PqqB. The finding of a protease that specifically targets serine side chains is rare, and we propose that this activity may be useful in proteomic analyses of the large family of proteins that have undergone post-translational phosphorylation at serine.
dc.description.sponsorshipNational Institutes of HealthUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USA [GM118117, GM124002, 1S10OD020062-01]
dc.identifier.doi10.1074/jbc.RA119.009684
dc.identifier.issn0021-9258
dc.identifier.urihttp://hdl.handle.net/10400.1/14219
dc.language.isoeng
dc.peerreviewedyes
dc.publisherAmerican Society for Biochemistry and Molecular Biology
dc.subjectGlucose-dehydrogenase
dc.subjectNucleotide-sequence
dc.subjectEscherichia-coli
dc.subjectMammalian enzyme
dc.subjectPqq
dc.subjectMethylobacterium
dc.subjectBiosynthesis
dc.subjectBiogenesis
dc.subjectSite
dc.subjectOxidation
dc.subjectProtease
dc.subjectZinc
dc.subjectSerine
dc.subjectEnzyme catalysis
dc.subjectPeptides
dc.subjectHeterodimer
dc.subjectM16B Protease
dc.subjectPQQ
dc.titleA two-component protease in Methylorubrum extorquens with high activity toward the peptide precursor of the redox cofactor pyrroloquinoline quinone
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage15036
oaire.citation.issue41
oaire.citation.startPage15025
oaire.citation.titleJournal of Biological Chemistry
oaire.citation.volume294
person.familyNameMartel
person.givenNamePaulo
person.identifier1955829
person.identifier.ciencia-idAD16-8C2A-26CE
person.identifier.orcid0000-0001-8237-1267
person.identifier.ridK-4704-2013
person.identifier.scopus-author-id7101625192
rcaap.rightsopenAccess
rcaap.typearticle
relation.isAuthorOfPublicatione70c3a99-3cde-4b3d-8ca2-4ac252906ae3
relation.isAuthorOfPublication.latestForDiscoverye70c3a99-3cde-4b3d-8ca2-4ac252906ae3

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