title: Analysis of JmjC Demethylase-Catalyzed Demethylation Using Geometrically-Constrained Lysine Analogues.
abstract: The dynamic post-translational modifications of histones play important roles in the regulation of transcription in animals. The demethylation of N(ε)-methyl lysine residues in the N-terminal tail of histone H3 is catalyzed by demethylases, of which the largest family is the ferrous iron and 2-oxoglutarate dependent demethylases (JmjC KDMs), which catalyze demethylation via initial hydroxylation of the N-methyl groups. We report studies on the conformational requirements of the JmjC KDM substrates using N-methylated lysine analogues prepared by metathesis reactions of suitably protected N-allylglycine. The results support the proposed requirement for a positively charged N(ε)-amino group in JmjC KDM catalysis. Demethylation of a trans-C-4/C-5 dehydrolysine substrate analogue was observed with representative KDM4 subfamily members KDM4A, KDM4B and KDM4E, and KDM7B, which are predicted, based on crystallographic analyses, to bind the N(ε)-methylated lysine residue in different conformations during catalysis. This information may be useful in the design of JmjC KDM selective inhibitors.
authors.affiliation: Chemistry Research Laboratory , 12 Mansfield Road, Oxford, OX1 3TA, United Kingdom.
authors.forename: Gareth W
authors.initials: GW
authors.lastname: Langley
authors.affiliation: Chemistry Research Laboratory , 12 Mansfield Road, Oxford, OX1 3TA, United Kingdom.|Department of Chemistry, Aarhus University , Langelandsgade 140, 8000 Aarhus C, Denmark.
authors.forename: Anne
authors.initials: A
authors.lastname: Brinkø
authors.affiliation: Chemistry Research Laboratory , 12 Mansfield Road, Oxford, OX1 3TA, United Kingdom.
authors.forename: Martin
authors.initials: M
authors.lastname: Münzel
authors.affiliation: Chemistry Research Laboratory , 12 Mansfield Road, Oxford, OX1 3TA, United Kingdom.
authors.forename: Louise J
authors.initials: LJ
authors.lastname: Walport
authors.affiliation: Chemistry Research Laboratory , 12 Mansfield Road, Oxford, OX1 3TA, United Kingdom.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Schofield
authors.affiliation: Chemistry Research Laboratory , 12 Mansfield Road, Oxford, OX1 3TA, United Kingdom.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Hopkinson
mesh_terms: D001665:Binding Sites; D002384:Catalysis; D056484:Jumonji Domain-Containing Histone Demethylases / Q000235:genetics / Q000378:metabolism*; D008239:Lysine / Q000031:analogs & derivatives* / Q000737:chemistry; D008745:Methylation; D013379:Substrate Specificity
chemical_list: D056484:Jumonji Domain-Containing Histone Demethylases; D008239:Lysine