title: Mechanism of N-Acylthiourea-mediated activation of human histone deacetylase 8 (HDAC8) at molecular and cellular levels.
abstract: We reported previously that an N-acylthiourea derivative (TM-2-51) serves as a potent and isozyme-selective activator for human histone deacetylase 8 (HDAC8). To probe the molecular mechanism of the enzyme activation, we performed a detailed account of the steady-state kinetics, thermodynamics, molecular modeling, and cell biology studies. The steady-state kinetic data revealed that TM-2-51 binds to HDAC8 at two sites in a positive cooperative manner. Isothermal titration calorimetric and molecular modeling data conformed to the two-site binding model of the enzyme-activator complex. We evaluated the efficacy of TM-2-51 on SH-SY5Y and BE(2)-C neuroblastoma cells, wherein the HDAC8 expression has been correlated with cellular malignancy. Whereas TM-2-51 selectively induced cell growth inhibition and apoptosis in SH-SY5Y cells, it showed no such effects in BE(2)-C cells, and this discriminatory feature appears to be encoded in the p53 genotype of the above cells. Our mechanistic and cellular studies on HDAC8 activation have the potential to provide insight into the development of novel anticancer drugs.
authors.affiliation: From the Departments of Chemistry and Biochemistry.
authors.forename: Raushan K
authors.initials: RK
authors.lastname: Singh
authors.affiliation: Animal Science.
authors.forename: Kyongshin
authors.initials: K
authors.lastname: Cho
authors.affiliation: Pharmaceutical Sciences, and.
authors.forename: Satish K R
authors.initials: SK
authors.lastname: Padi
authors.affiliation: From the Departments of Chemistry and Biochemistry.
authors.forename: Junru
authors.initials: J
authors.lastname: Yu
authors.affiliation: Pharmaceutical Sciences, and.
authors.forename: Manas
authors.initials: M
authors.lastname: Haldar
authors.affiliation: From the Departments of Chemistry and Biochemistry.
authors.forename: Tanmay
authors.initials: T
authors.lastname: Mandal
authors.affiliation: Computer Science, North Dakota State University, Fargo, North Dakota 58108.
authors.forename: Changhui
authors.initials: C
authors.lastname: Yan
authors.affiliation: From the Departments of Chemistry and Biochemistry.
authors.forename: Gregory
authors.initials: G
authors.lastname: Cook
authors.affiliation: Pharmaceutical Sciences, and.
authors.forename: Bin
authors.initials: B
authors.lastname: Guo
authors.affiliation: Pharmaceutical Sciences, and.
authors.forename: Sanku
authors.initials: S
authors.lastname: Mallik
authors.affiliation: From the Departments of Chemistry and Biochemistry, dk.srivastava@ndsu.edu.
authors.forename: D K
authors.initials: DK
authors.lastname: Srivastava
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D001549:Benzamides / Q000008:administration & dosage; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D018360:Crystallography, X-Ray; D004789:Enzyme Activation / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic; D056572:Histone Deacetylase Inhibitors / Q000008:administration & dosage; D006655:Histone Deacetylases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D009447:Neuroblastoma / Q000188:drug therapy / Q000201:enzymology* / Q000235:genetics; D010670:Phenylthiourea / Q000008:administration & dosage / Q000031:analogs & derivatives; D012097:Repressor Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D013816:Thermodynamics; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis
chemical_list: D001549:Benzamides; D056572:Histone Deacetylase Inhibitors; C571363:N-(phenylcarbothiol)benzamide; D012097:Repressor Proteins; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D010670:Phenylthiourea; C409470:HDAC8 protein, human; D006655:Histone Deacetylases
keywords: Calorimetry; Catalysis; Histone Deacetylase (HDAC); Neuroblastoma; p53