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Title: 1,2-Propanediol Dehydration in Roseburia inulinivorans: Structural Basis for Substrate and Enantiomer Selectivity

Journal Article · · Journal of Biological Chemistry

Glycyl radical enzymes (GREs) represent a diverse superfamily of enzymes that utilize a radical mechanism to catalyze difficult, but often essential, chemical reactions. Here, we present the first biochemical and structural data for a GRE-type diol dehydratase from the organism Roseburia inulinivorans (RiDD). Despite high sequence (48% identity) and structural similarity to the GRE-type glycerol dehydratase from Clostridium butyricum, we demonstrate that the RiDD is in fact a diol dehydratase. In addition, the RiDD will utilize both (S)-1,2- propanediol and (R)-1,2-propanediol as a substrate, with an observed preference for the S enantiomer. Based on the new structural information we developed and successfully tested a hypothesis that explains the functional differences we observe.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Organization:
USDOE
OSTI ID:
1324768
Journal Information:
Journal of Biological Chemistry, Vol. 291, Issue 30; ISSN 0021-9258
Publisher:
American Society for Biochemistry and Molecular BiologyCopyright Statement
Country of Publication:
United States
Language:
ENGLISH
Citation Metrics:
Cited by: 23 works
Citation information provided by
Web of Science

References (55)

Pyruvate formate-lyase mechanism involving the protein-based glycyl radical journal August 1993
Insights into the Glycyl Radical Enzyme Active Site of Benzylsuccinate Synthase: A Computational Study journal August 2013
Crystal Structure of E.coli Alcohol Dehydrogenase YqhD: Evidence of a Covalently Modified NADP Coenzyme journal September 2004
p -Hydroxyphenylacetate decarboxylase from Clostridium difficile : A novel glycyl radical enzyme catalysing the formation of journal March 2001
Re-evaluation of EDTA-Chelated Biuret Reagent journal October 1974
Enzymatic Radical Catalysis: Coenzyme B12-Dependent Diol Dehydratase journal September 2002
Structures of benzylsuccinate synthase elucidate roles of accessory subunits in glycyl radical enzyme activation and activity journal June 2014
Radical SAM Activation of the B 12 -Independent Glycerol Dehydratase Results in Formation of 5′-Deoxy-5′-(methylthio)adenosine and Not 5′-Deoxyadenosine journal February 2011
Glycyl radical activating enzymes: Structure, mechanism, and substrate interactions journal March 2014
The [4Fe-4S] 1+ Cluster of Pyruvate Formate-Lyase Activating Enzyme Generates the Glycyl Radical on Pyruvate Formate-Lyase:  EPR-Detected Single Turnover journal August 2000
GC Determination of Acetone, Acetaldehyde, Ethanol, and Methanol in Biological Matrices and Cell Culture journal April 2009
The Free Radical of the Anaerobic Ribonucleotide Reductase from Escherichia coli Is at Glycine 681 journal March 1996
Size-Distribution Analysis of Macromolecules by Sedimentation Velocity Ultracentrifugation and Lamm Equation Modeling journal March 2000
Subunit Structure of Benzylsuccinate Synthase journal February 2009
Inference of Macromolecular Assemblies from Crystalline State journal September 2007
Microbial conversion of choline to trimethylamine requires a glycyl radical enzyme journal November 2012
Inactivation of E. coli pyruvate formate-lyase: Role of AdhE and small molecules journal March 2007
A Stable Organic Free Radical in Anaerobic Benzylsuccinate Synthase of Azoarcus sp. Strain T journal January 2001
A radical-chemical route to acetyl-CoA: the anaerobically induced pyruvate formate-lyase system of Escherichia coli journal August 1990
Structural Rationalization for the Lack of Stereospecificity in Coenzyme B 12 -dependent Diol Dehydratase journal April 2003
The Stability of C α Peptide Radicals: Why Glycyl Radical Enzymes? journal February 2011
Influence of Tris(Hydroxymethyl)Aminomethane on Kinetic Mechanism of Yeast Alcohol Dehydrogenase journal January 1998
Radical Mechanisms of Enzymatic Catalysis journal June 2001
In Vitro Characterization of AtsB, a Radical SAM Formylglycine-Generating Enzyme That Contains Three [4Fe-4S] Clusters journal July 2008
Coot model-building tools for molecular graphics journal November 2004
Structure and Function of CutC Choline Lyase from Human Microbiota Bacterium Klebsiella pneumoniae journal July 2015
Molecular characterization of the 1,3-propanediol (1,3-PD) operon of Clostridium butyricum journal April 2003
[23] Glycyl free radical in pyruvate formate-lyase: Synthesis, structure characteristics, and involvement in catalysis book January 1995
Structural Basis for a Kolbe-Type Decarboxylation Catalyzed by a Glycyl Radical Enzyme journal September 2011
Characterization of Choline Trimethylamine-Lyase Expands the Chemistry of Glycyl Radical Enzymes journal May 2014
PHENIX: a comprehensive Python-based system for macromolecular structure solution journal January 2010
Generation of the Glycyl Radical of the Anaerobic Escherichia coli Ribonucleotide Reductase Requires a Specific Activating Enzyme journal February 1995
Catalytic Mechanism of the Glycyl Radical Enzyme 4-Hydroxyphenylacetate Decarboxylase from Continuum Electrostatic and QC/MM Calculations journal September 2013
Chemical and Biological Reduction of the Radical SAM Enzyme CPH 4 Synthase journal April 2015
Pyruvate Formate-lyase and Its Activation by Pyruvate Formate-lyase Activating Enzyme journal December 2013
Redesign of coenzyme B12 dependent diol dehydratase to be resistant to the mechanism-based inactivation by glycerol and act on longer chain 1,2-diols: Redesign of B12 dependent diol dehydratase journal January 2012
Purification and crystallization of a proteolytic fragment of Escherichia coli pyruvate formate-lyase journal February 1999
Free R value: a novel statistical quantity for assessing the accuracy of crystal structures journal January 1992
A Glycyl Radical Site in the Crystal Structure of a Class III Ribonucleotide Reductase journal March 1999
Expression of 1,3-propanediol oxidoreductase and its isoenzyme in Klebsiella pneumoniae for bioconversion of glycerol into 1,3-propanediol journal May 2010
Implications of the Inability of Listeria monocytogenes EGD-e To Grow Anaerobically Due to a Deletion in the Class III NrdD Ribonucleotide Reductase for Its Use as a Model Laboratory Strain journal April 2011
Insight into the Mechanism of the B 12 -Independent Glycerol Dehydratase from Clostridium butyricum :  Preliminary Biochemical and Structural Characterization journal April 2004
Prediction of Hydrodynamic and Other Solution Properties of Rigid Proteins from Atomic- and Residue-Level Models journal August 2011
Glycerol Dehydratation by the B 12 -Independent Enzyme May Not Involve the Migration of a Hydroxyl Group: A Computational Study journal June 2012
Reconstitution and Characterization of the Polynuclear Iron-Sulfur Cluster in Pyruvate Formate-lyase-activating Enzyme: MOLECULAR PROPERTIES OF THE HOLOENZYME FORM journal February 1998
YqhD: a broad-substrate range aldehyde reductase with various applications in production of biorenewable fuels and chemicals journal October 2010
Adenosylmethionine-dependent synthesis of the glycyl radical in pyruvate formate-lyase by abstraction of the glycine C-2 pro-S hydrogen atom. Studies of [2H]glycine-substituted enzyme and peptides homologous to the glycine 734 site. journal April 1994
Purification of 1,3-propanediol dehydrogenase from Citrobacter freundii and cloning, sequencing, and overexpression of the corresponding gene in Escherichia coli. journal January 1995
Cavin1 intrinsically disordered domains are essential for fuzzy electrostatic interactions and caveola formation journal February 2021
PHENIX: a comprehensive Python-based system for macromolecular structure solution. text January 2010
A spectrophotometric method for the direct recording of dioldehydrase activity journal October 1971
A radical-chemical route to acetyl-CoA: the anaerobically induced pyruvate formate-lyase system of journal August 1990
Pyruvate formate lyase is structurally homologous to type I ribonucleotide reductase journal July 1999
Structure of Escherichia coli pyruvate formate-lyase with pyruvate journal November 2002
New glycyl radical enzymes catalysing key metabolic steps in anaerobic bacteria journal January 2005

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