title: Correlation of structure and function in the human hotdog-fold enzyme hTHEM4.
abstract: Human THEM4 (hTHEM4) is comprised of a catalytically active hotdog-fold acyl-CoA thioesterase domain and an N-terminal domain of unknown fold and function. hTHEM4 has been linked to Akt1 regulation and cell apoptosis. Herein, we report the X-ray structure of hHTEM4 bound with undecan-2-one-CoA. Structure guided mutagenesis was carried out to confirm the catalytic residues. The N-terminal domain is shown to be partially comprised of irregular and flexible secondary structure, reminiscent of a protein-binding domain. We demonstrate direct hTHEM4-Akt1 binding by immunoprecipitation and by inhibition of Akt1 kinase activity, thus providing independent evidence that hTHEM4 is an Akt1 negative regulator.
authors.affiliation: Department of Chemistry and Chemical Biology, University of New Mexico, Albuquerque, NM 87131, USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Zhao
authors.forename: Kap
authors.initials: K
authors.lastname: Lim
authors.forename: Anthony
authors.initials: A
authors.lastname: Choudry
authors.forename: John A
authors.initials: JA
authors.lastname: Latham
authors.forename: Manish C
authors.initials: MC
authors.lastname: Pathak
authors.forename: Dennis
authors.initials: D
authors.lastname: Dominguez
authors.forename: Lusong
authors.initials: L
authors.lastname: Luo
authors.forename: Osnat
authors.initials: O
authors.lastname: Herzberg
authors.forename: Debra
authors.initials: D
authors.lastname: Dunaway-Mariano
mesh_terms: D000214:Acyl Coenzyme A / Q000737:chemistry / Q000378:metabolism; D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry* / Q000378:metabolism; D020134:Catalytic Domain / Q000187:drug effects; D018360:Crystallography, X-Ray; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D017433:Protein Structure, Secondary; D051057:Proto-Oncogene Proteins c-akt / Q000737:chemistry / Q000378:metabolism; D013869:Thiolester Hydrolases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism
chemical_list: D000214:Acyl Coenzyme A; D048868:Adaptor Proteins, Signal Transducing; D008565:Membrane Proteins; D051057:Proto-Oncogene Proteins c-akt; C437985:THEM4 protein, human; D013869:Thiolester Hydrolases