Structural and biochemical characterization of EDTA monooxygenase and its physical interaction with a partner flavin reductase.


:Ethylenediaminetetraacetate (EDTA) is currently the most abundant organic pollutant due to its recalcitrance and extensive use. Only a few bacteria can degrade it, using EDTA monooxygenase (EmoA) to initiate the degradation. EmoA is an FMNH2 -dependent monooxygenase that requires an NADH:FMN oxidoreductase (EmoB) to provide FMNH2 as a cosubstrate. Although EmoA has been identified from Chelativorans (ex. Mesorhizobium) sp. BNC1, its catalytic mechanism is unknown. Crystal structures of EmoA revealed a domain-like insertion into a TIM-barrel, which might serve as a flexible lid for the active site. Docking of MgEDTA(2-) into EmoA identified an intricate hydrogen bond network connected to Tyr(71) , which should potentially lower its pKa. Tyr(71) , along with nearby Glu(70) and a peroxy flavin, facilitates a keto-enol transition of the leaving acetyl group of EDTA. Further, for the first time, the physical interaction between EmoA and EmoB was observed by ITC, molecular docking and enzyme kinetic assay, which enhanced both EmoA and EmoB activities probably through coupled channelling of FMNH2 .


Mol Microbiol


Molecular microbiology


Jun SY,Lewis KM,Youn B,Xun L,Kang C




Has Abstract


2016-06-01 00:00:00












  • N-mediated transcription antitermination in lambdoid phage H-19B is characterized by alternative NUT RNA structures and a reduced requirement for host factors.

    abstract::Gene expression in lambdoid phages in part is controlled by transcription antitermination. For most lambdoid phages, maximal expression of delayed early genes requires an RNA polymerase modified by the phage N and host Nus proteins at RNA NUT sites. The NUT sites (NUTL and NUTR) are made up of three elements: BOXA, BO...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Neely MN,Friedman DI

    更新日期:2000-12-01 00:00:00

  • Streptococcus pyogenes antigen I/II-family polypeptide AspA shows differential ligand-binding properties and mediates biofilm formation.

    abstract::The streptococcal antigen I/II (AgI/II)-family polypeptides are cell wall-anchored adhesins expressed by most indigenous oral streptococci. Proteins sharing 30-40% overall amino acid sequence similarities with AgI/II-family proteins are also expressed by Streptococcus pyogenes. The S. pyogenes M28_Spy1325 polypeptide ...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Maddocks SE,Wright CJ,Nobbs AH,Brittan JL,Franklin L,Strömberg N,Kadioglu A,Jepson MA,Jenkinson HF

    更新日期:2011-08-01 00:00:00

  • Pkn9, a Ser/Thr protein kinase involved in the development of Myxococcus xanthus.

    abstract::The Myxococcus xanthus gene, pkn9, encodes a protein that contains significant homology with eukaryotic Ser/Thr protein kinases. The pkn9 gene was singled out of a previously identified family of kinase genes by amplification techniques that displayed differences in kinase gene expression during selected periods of th...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Hanlon WA,Inouye M,Inouye S

    更新日期:1997-02-01 00:00:00

  • Plant defence suppression is mediated by a fungal sirtuin during rice infection by Magnaporthe oryzae.

    abstract::Crop destruction by the hemibiotrophic rice pathogen Magnaporthe oryzae requires plant defence suppression to facilitate extensive biotrophic growth in host cells before the onset of necrosis. How this is achieved at the genetic level is not well understood. Here, we report that a M. oryzae sirtuin, MoSir2, plays an e...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Fernandez J,Marroquin-Guzman M,Nandakumar R,Shijo S,Cornwell KM,Li G,Wilson RA

    更新日期:2014-10-01 00:00:00

  • An alternative PII protein in the regulation of glutamine synthetase in Escherichia coli.

    abstract::The PII protein has been considered pivotal to the dual cascade regulating ammonia assimilation through glutamine synthetase activity. Here we show that PII, encoded by the glnB gene, is not always essential; for instance upon ammonia deprivation of a glnB deletion strain, glutamine synthetase can be deadenylylated as...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: van Heeswijk WC,Hoving S,Molenaar D,Stegeman B,Kahn D,Westerhoff HV

    更新日期:1996-07-01 00:00:00

  • Families of bacterial signal-transducing proteins.

    abstract::Bacteria can respond to a variety of environmental stimuli by means of systems generally composed of two proteins. The first protein (sensor or transmitter) is usually a transmembrane protein with cytoplasmic and extracytoplasmic domains. The extracytoplasmic domain (sensor) senses the environment and transfers the si...

    journal_title:Molecular microbiology

    pub_type: 杂志文章,评审


    authors: Gross R,Aricò B,Rappuoli R

    更新日期:1989-11-01 00:00:00

  • YscP, a Yersinia protein required for Yop secretion that is surface exposed, and released in low Ca2+.

    abstract::The Yersinia Ysc apparatus is made of more than 20 proteins, 11 of which have homologues in many type III systems. Here, we characterize YscP from Yersinia enterocolitica. This 515-residue protein has a high proline content, a large tandem repetition and a slow migration in SDS-PAGE. Unlike the products of neighbourin...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Stainier I,Bleves S,Josenhans C,Karmani L,Kerbourch C,Lambermont I,Tötemeyer S,Boyd A,Cornelis GR

    更新日期:2000-09-01 00:00:00

  • Use of rare-earth elements in the phyllosphere colonizer Methylobacterium extorquens PA1.

    abstract::Until recently, rare-earth elements (REEs) had been thought to be biologically inactive. This view changed with the discovery of the methanol dehydrogenase XoxF that strictly relies on REEs for its activity. Some methylotrophs only contain xoxF, while others, including the model phyllosphere colonizer Methylobacterium...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Ochsner AM,Hemmerle L,Vonderach T,Nüssli R,Bortfeld-Miller M,Hattendorf B,Vorholt JA

    更新日期:2019-05-01 00:00:00

  • Structural, genetic and functional characterization of the flagellin glycosylation process in Helicobacter pylori.

    abstract::Mass spectrometry analyses of the complex polar flagella from Helicobacter pylori demonstrated that both FlaA and FlaB proteins are post-translationally modified with pseudaminic acid (Pse5Ac7Ac, 5,7-diacetamido-3,5,7,9-tetradeoxy-l-glycero-l-manno -n o n-ulosonic acid). Unlike Campylobacter, flagellar glycosylation i...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Schirm M,Soo EC,Aubry AJ,Austin J,Thibault P,Logan SM

    更新日期:2003-06-01 00:00:00

  • Escherichia coli translocase: the unravelling of a molecular machine.

    abstract::Protein translocation across the bacterial cytoplasmic membrane has been studied extensively in Escherichia coli. The identification of the components involved and subsequent reconstitution of the purified translocation reaction have defined the minimal constituents that allowed extensive biochemical characterization ...

    journal_title:Molecular microbiology

    pub_type: 杂志文章,评审


    authors: Manting EH,Driessen AJ

    更新日期:2000-07-01 00:00:00

  • Eeyarestatin 24 impairs SecYEG-dependent protein trafficking and inhibits growth of clinically relevant pathogens.

    abstract::Eeyarestatin 1 (ES1) is an inhibitor of endoplasmic reticulum (ER) associated protein degradation, Sec61-dependent Ca2+ homeostasis and protein translocation into the ER. Recently, evidence was presented showing that a smaller analog of ES1, ES24, targets the Sec61-translocon, and captures it in an open conformation t...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Steenhuis M,Koningstein GM,Oswald J,Pick T,O'Keefe S,Koch HG,Cavalié A,Whitehead RC,Swanton E,High S,Luirink J

    更新日期:2020-08-15 00:00:00

  • UvrD-dependent replication of rolling-circle plasmids in Escherichia coli.

    abstract::The Escherichia coli UvrD helicase (or helicase II) is known for its involvement in DNA repair. We report that UvrD is required for DNA replication of several different rolling-circle plasmids in E. coli, whereas its homologue, the Rep helicase, is not. Lack of UvrD helicase does not impair the first step of plasmid r...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Bruand C,Ehrlich SD

    更新日期:2000-01-01 00:00:00

  • Tandem DNA-bound cAMP-CRP complexes are required for transcriptional repression of the deoP2 promoter by the CytR repressor in Escherichia coli.

    abstract::We have studied the deoP2 promoter in Escherichia coli to define features important for its interaction with the CytR repressor. As is characteristic for CytR-regulated promoters, deoP2 encodes tandem binding sites for the activating complex cAMP-CRP. One of these sites, CRP-1, overlaps the - 35 region, and is suffici...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Sogaard-Andersen L,Mellegaard NE,Douthwaite SR,Valentin-Hansen P

    更新日期:1990-09-01 00:00:00

  • Structural mechanisms of multidrug recognition and regulation by bacterial multidrug transcription factors.

    abstract::The increase in bacterial resistance to multiple drugs represents a serious and growing health risk. One component of multidrug resistance (MDR) is a group of multidrug transporters that are often regulated at the transcriptional level by repressors and/or activators. Some of these transcription factors are also multi...

    journal_title:Molecular microbiology

    pub_type: 杂志文章,评审


    authors: Schumacher MA,Brennan RG

    更新日期:2002-08-01 00:00:00

  • Helicobacter pylori HP0518 affects flagellin glycosylation to alter bacterial motility.

    abstract::Helicobacter pylori is a human gastric pathogen associated with gastric and duodenal ulcers as well as gastric cancer. Mounting evidence suggests this pathogen's motility is prerequisite for successful colonization of human gastric tissues. Here, we isolated an H. pylori G27 HP0518 mutant exhibiting altered motility i...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Asakura H,Churin Y,Bauer B,Boettcher JP,Bartfeld S,Hashii N,Kawasaki N,Mollenkopf HJ,Jungblut PR,Brinkmann V,Meyer TF

    更新日期:2010-12-01 00:00:00

  • Post-transcriptional regulation by distal Shine-Dalgarno sequences in the grpE-dnaK intergenic region of Streptococcus mutans.

    abstract::A unique 373 bp region (igr66) between grpE and dnaK of Streptococcus mutans lacks a promoter but is required for optimal production of DnaK. Northern blotting using probes specific to hrcA, igr66 or dnaK revealed multiple transcripts produced from the dnaK operon and 5'-RACE mapped 5' termini of multiple dnaK transcr...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Palmer SR,Burne RA

    更新日期:2015-10-01 00:00:00

  • Identification of structural and molecular determinants of the tyrosine-kinase Wzc and implications in capsular polysaccharide export.

    abstract::Capsular polysaccharides are well-established virulence factors of pathogenic bacteria. Their biosynthesis and export are regulated within the transmembrane polysaccharide assembly machinery by the autophosphorylation of atypical tyrosine-kinases, named BY-kinases. However, the accurate functioning of these tyrosine-k...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Bechet E,Gruszczyk J,Terreux R,Gueguen-Chaignon V,Vigouroux A,Obadia B,Cozzone AJ,Nessler S,Grangeasse C

    更新日期:2010-09-01 00:00:00

  • Gene expression in mycobacteria: transcriptional fusions based on xylE and analysis of the promoter region of the response regulator mtrA from Mycobacterium tuberculosis.

    abstract::Understanding promoter regulation and signal-transduction systems in pathogenic mycobacteria is critical for uncovering the processes that govern interactions of these bacteria with the human host. In order to develop additional genetic tools for analysis of mycobacterial promoters, the xyIE gene from Pseudomonas was ...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Curcic R,Dhandayuthapani S,Deretic V

    更新日期:1994-09-01 00:00:00

  • A fresh view of the cell biology of copper in enterobacteria.

    abstract::Copper ions are essential but also very toxic. Copper resistance in bacteria is based on export of the toxic ion, oxidation from Cu(I) to Cu(II), and sequestration by copper-binding metal chaperones, which deliver copper ions to efflux systems or metal-binding sites of copper-requiring proteins. In their publication i...

    journal_title:Molecular microbiology

    pub_type: 评论,杂志文章


    authors: Nies DH,Herzberg M

    更新日期:2013-02-01 00:00:00

  • Listeria monocytogenes virulence proteins induce surface expression of Fas ligand on T lymphocytes.

    abstract::Virulence factors secreted by Listeria monocytogenes are known to interfere with host cellular signalling pathways. We investigated whether L. monocytogenes modulates T-cell receptor signalling by examining surface expression of proteins known to be upregulated on activated T cells. In vitro culture of murine splenocy...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Zenewicz LA,Skinner JA,Goldfine H,Shen H

    更新日期:2004-03-01 00:00:00

  • Microtubule-severing proteins are involved in flagellar length control and mitosis in Trypanosomatids.

    abstract::Microtubules are key players in the biology of Trypanosomatid parasites, not only as classical components of the mitotic spindle, microtubule-organizing centres and flagellum but also as the essential constituent of the cytoskeleton. Their length dynamics are regulated by, among others, microtubule-severing proteins. ...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Casanova M,Crobu L,Blaineau C,Bourgeois N,Bastien P,Pagès M

    更新日期:2009-03-01 00:00:00

  • Hyperosmosis enhances radiation and hydroxyurea resistance of Schizosaccharomyces pombe checkpoint mutants through the spindle checkpoint and delayed cytokinesis.

    abstract::The DNA damage and stress response pathways interact to regulate cellular responses to genotoxins and environmental stresses. How these pathways interact in Schizosaccharomyces pombe is not well understood. We demonstrate that osmotic stress suppresses the DNA damage sensitivity of checkpoint mutants, and that this oc...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Alao JP,Huis In 't Veld PJ,Buhse F,Sunnerhagen P

    更新日期:2010-07-01 00:00:00

  • Structure and function of DnaA and the DnaA-box in eubacteria: evolutionary relationships of bacterial replication origins.

    abstract::DnaA protein (a trans-acting element) and its binding sequence, DnaA-box: (a cis-acting element) are two elements essential for the initiation of chromosomal replication in Escherichia coli and other enteric bacteria. Recently these two elements have been found to be conserved in three Gram-positive bacteria (Bacillus...

    journal_title:Molecular microbiology

    pub_type: 杂志文章,评审


    authors: Yoshikawa H,Ogasawara N

    更新日期:1991-11-01 00:00:00

  • Molecular cloning, partial purification, and characterization of a haemin-binding lipoprotein from Haemophilus influenzae type b.

    abstract::A library of genomic DNA fragments from Haemophilus influenzae type b (Hib) DL42 was constructed in plasmid pBR322, transformed into Escherichia coli strain RR1, and screened for recombinant clones with haemin-binding activity by plating onto haemin-containing agar. Expression of haemin-binding activity by clones corr...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Hanson MS,Hansen EJ

    更新日期:1991-02-01 00:00:00

  • DNA replication-coupled inactivation of DnaA protein in vitro: a role for DnaA arginine-334 of the AAA+ Box VIII motif in ATP hydrolysis.

    abstract::The DnaA protein, which initiates chromosomal replication in Escherichia coli, is negatively regulated by both the sliding clamp of DNA polymerase III holoenzyme and the IdaB protein. We have found that, when the amount of minichromosome is limited in an in vitro replication system, minichromosomal replication-stimula...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Su'etsugu M,Kawakami H,Kurokawa K,Kubota T,Takata M,Katayama T

    更新日期:2001-04-01 00:00:00

  • Roles for both FtsA and the FtsBLQ subcomplex in FtsN-stimulated cell constriction in Escherichia coli.

    abstract::Escherichia coli FtsN is a bitopic membrane protein that is essential for triggering active cell constriction. A small periplasmic subdomain ((E) FtsN) is required and sufficient for function, but its mechanism of action is unclear. We isolated extragenic (E) FtsN*-suppressing mutations that restore division in cells ...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Liu B,Persons L,Lee L,de Boer PA

    更新日期:2015-03-01 00:00:00

  • In vitro reconstitution of a trimeric complex of DivIB, DivIC and FtsL, and their transient co-localization at the division site in Streptococcus pneumoniae.

    abstract::DivIB, DivIC and FtsL are bacterial proteins essential for cell division, which show interdependencies for their stabilities and localization. We have reconstituted in vitro a trimeric complex consisting of the recombinant extracellular domains of the three proteins from Streptococcus pneumoniae. The extracellular dom...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Noirclerc-Savoye M,Le Gouëllec A,Morlot C,Dideberg O,Vernet T,Zapun A

    更新日期:2005-01-01 00:00:00

  • Septal membrane fusion--a pivotal event in bacterial spore formation?

    abstract::Formation of the asymmetrically located septum divides sporulating bacilli into two distinct cells: the mother cell and the prespore. The rigidifying wall material in the septum is subsequently removed by autolysis. Examination of published electron micrographs indicates that the two septal membranes then fuse to form...

    journal_title:Molecular microbiology

    pub_type: 杂志文章,评审


    authors: Higgins ML,Piggot PJ

    更新日期:1992-09-01 00:00:00

  • New mechanistic insights into pre-protein transport across the second outermost plastid membrane of diatoms.

    abstract::Chromalveolates like the diatom Phaeodactylum tricornutum arose through the uptake of a red alga by a phagotrophic protist, a process termed secondary endosymbiosis. In consequence, the plastids are surrounded by two additional membranes compared with primary plastids. This plastid morphology poses additional barriers...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Hempel F,Felsner G,Maier UG

    更新日期:2010-05-01 00:00:00

  • Expression of the Escherichia coli pcnB gene is translationally limited using an inefficient start codon: a second chromosomal example of translation initiated at AUU.

    abstract::Expression of the gene pcnB, encoding the dispensable Escherichia coli poly(A) polymerase (PAPI), which is toxic when overproduced, was investigated. Its promoter was identified and found to be moderately strong when used to express a beta-galactosidase reporter. Expression levels were not affected by increasing or de...

    journal_title:Molecular microbiology

    pub_type: 杂志文章


    authors: Binns N,Masters M

    更新日期:2002-06-01 00:00:00