title: 1H,15N,13C backbone resonance assignments of human soluble catechol O-methyltransferase in complex with S-adenosyl-L-methionine and 3,5-dinitrocatechol.
abstract: Catechol O-methyltransferase (COMT) is an enzyme that plays a major role in catechol neurotransmitter deactivation. Inhibition of COMT can increase neurotransmitter levels, which provides a means of treatment for Parkinson's disease, schizophrenia and depression. COMT exists as two isozymes: a soluble cytoplasmic form (S-COMT), expressed in the liver and kidneys and a membrane-bound form (MB-COMT), found mostly in the brain. Here we report the backbone1H,15N and13C chemical shift assignments of S-COMT in complex with S-adenosyl-L-methionine, 3,5-dinitrocatechol and Mg2+. Assignments were obtained by heteronuclear multidimensional NMR spectroscopy. In total, 97 % of all backbone resonances were assigned in the complex, with 205 out of a possible 215 residues assigned in the1H-15N TROSY spectrum. Prediction of solution secondary structure from a chemical shift analysis using the TALOS+ webserver is in good agreement with published X-ray crystal structures.
authors.affiliation: Manchester Institute of Biotechnology and School of Chemistry, The University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK.
authors.forename: Sylwia
authors.initials: S
authors.lastname: Czarnota
authors.affiliation: Manchester Institute of Biotechnology and School of Chemistry, The University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK. n.j.baxter@sheffield.ac.uk.|Department of Molecular Biology and Biotechnology, Krebs Institute for Biomolecular Research, The University of Sheffield, Firth Court, Western Bank, Sheffield, S10 2TN, UK. n.j.baxter@sheffield.ac.uk.
authors.forename: Nicola J
authors.initials: NJ
authors.lastname: Baxter
authors.affiliation: Manchester Institute of Biotechnology and School of Chemistry, The University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Cliff
authors.affiliation: Manchester Institute of Biotechnology and School of Chemistry, The University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK.|Department of Molecular Biology and Biotechnology, Krebs Institute for Biomolecular Research, The University of Sheffield, Firth Court, Western Bank, Sheffield, S10 2TN, UK.
authors.forename: Jonathan P
authors.initials: JP
authors.lastname: Waltho
authors.affiliation: Manchester Institute of Biotechnology and School of Chemistry, The University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK.
authors.forename: Nigel S
authors.initials: NS
authors.lastname: Scrutton
authors.affiliation: Manchester Institute of Biotechnology and School of Chemistry, The University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK. sam.hay@manchester.ac.uk.
authors.forename: Sam
authors.identifier: 0000-0003-3274-0938
authors.initials: S
authors.lastname: Hay
mesh_terms: D001665:Binding Sites; D002394:Catechol O-Methyltransferase / Q000737:chemistry* / Q000378:metabolism*; D002396:Catechols / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D012436:S-Adenosylmethionine / Q000378:metabolism*; D012995:Solubility
chemical_list: D002396:Catechols; D012436:S-Adenosylmethionine; D002394:Catechol O-Methyltransferase; C057726:OR486
keywords: Backbone resonance assignment; Enzyme; S-adenosyl-L-methionine; Transverse relaxation optimized spectroscopy; Triple-labelled Protein