title: Azobenzene switch with a long-lived cis-state to photocontrol the enzyme activity of a histone deacetylase-like amidohydrolase.
abstract: The control of enzymes by use of an external stimulus such as light enables the temporal and spatial regulation of defined chemical reactions in a highly precise manner. In this work we investigated and characterized the reversible photocontrol of a bacterial histone deacetylase-like amidohydrolase (HDAH) from Bordetella/Alcaligenes strain FB188, which holds great potential to control deacetylation reactions of a broad spectrum of substrates in biotechnological and biomedical applications. Several HDAH variants with a single surface accessible cysteine close to the active site were developed and covalently modified by a monofunctional azobenzene-based photoswitch [4-phenylazomaleinanil (4-PAM)]. The enzymatic activity of three HDAH variants (M30C, S20C and M150C) were shown to be controlled by light. The thermal cis-to-trans relaxation of azobenzene conjugated to HDAH was up to 50-fold retarded compared to unbound 4-PAM allowing light pulse switching rather than continuing irradiation to maintain the thermodynamically less stable cis-state of covalently attached 4-PAM.
authors.forename: Michael
authors.initials: M
authors.lastname: Korbus
authors.forename: Ganesh
authors.initials: G
authors.lastname: Balasubramanian
authors.forename: Florian
authors.initials: F
authors.lastname: Müller-Plathe
authors.forename: Harald
authors.initials: H
authors.lastname: Kolmar
authors.forename: Franz-Josef
authors.initials: FJ
authors.lastname: Meyer-Almes
mesh_terms: D000581:Amidohydrolases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001391:Azo Compounds / Q000737:chemistry* / Q000378:metabolism*; D001884:Bordetella / Q000201:enzymology; D018360:Crystallography, X-Ray; D004789:Enzyme Activation; D014644:Genetic Variation / Q000235:genetics; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D055668:Photochemical Processes; D013237:Stereoisomerism; D013696:Temperature
chemical_list: D001391:Azo Compounds; D000581:Amidohydrolases; C009850:azobenzene