title: Structure and Function of CutC Choline Lyase from Human Microbiota Bacterium Klebsiella pneumoniae.
abstract: CutC choline trimethylamine-lyase is an anaerobic bacterial glycyl radical enzyme (GRE) that cleaves choline to produce trimethylamine (TMA) and acetaldehyde. In humans, TMA is produced exclusively by the intestinal microbiota, and its metabolite, trimethylamine oxide, has been associated with a higher risk of cardiovascular diseases. Therefore, information about the three-dimensional structures of TMA-producing enzymes is important for microbiota-targeted drug discovery. We have cloned, expressed, and purified the CutC GRE and the activating enzyme CutD from Klebsiella pneumoniae, a representative of the human microbiota. We have determined the first crystal structures of both the choline-bound and choline-free forms of CutC and have discovered that binding of choline at the ligand-binding site triggers conformational changes in the enzyme structure, a feature that has not been observed for any other characterized GRE.
authors.affiliation: From the Latvian Biomedical Research and Study Center, LV-1067 Riga, gints@biomed.lu.lv.
authors.forename: Gints
authors.initials: G
authors.lastname: Kalnins
authors.affiliation: the Latvian Institute of Organic Synthesis, LV-1006 Riga, and.
authors.forename: Janis
authors.initials: J
authors.lastname: Kuka
authors.affiliation: the Latvian Institute of Organic Synthesis, LV-1006 Riga, and.
authors.forename: Solveiga
authors.initials: S
authors.lastname: Grinberga
authors.affiliation: the Latvian Institute of Organic Synthesis, LV-1006 Riga, and.
authors.forename: Marina
authors.initials: M
authors.lastname: Makrecka-Kuka
authors.affiliation: the Latvian Institute of Organic Synthesis, LV-1006 Riga, and.
authors.forename: Edgars
authors.initials: E
authors.lastname: Liepinsh
authors.affiliation: the Latvian Institute of Organic Synthesis, LV-1006 Riga, and.
authors.forename: Maija
authors.initials: M
authors.lastname: Dambrova
authors.affiliation: From the Latvian Biomedical Research and Study Center, LV-1067 Riga, the University of Latvia, LV-1586 Riga, Latvia.
authors.forename: Kaspars
authors.initials: K
authors.lastname: Tars
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D020134:Catalytic Domain; D002794:Choline / Q000378:metabolism*; D002850:Chromatography, Gel; D002918:Chymotrypsin / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007711:Klebsiella pneumoniae / Q000201:enzymology*; D008190:Lyases / Q000737:chemistry* / Q000378:metabolism*; D064307:Microbiota; D008958:Models, Molecular; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D001426:Bacterial Proteins; D002918:Chymotrypsin; D008190:Lyases; D002794:Choline
keywords: choline; conformational change; enzyme catalysis; enzyme structure; radical