Department of Chemistry and Chemical Biology, University of New Mexico, Albuquerque, NM 87131, USA.
Biochemistry. 2012 Aug 21;51(33):6490-2. doi: 10.1021/bi300968n. Epub 2012 Aug 9.
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.
人 THEM4（hTHEM4）由一个催化活性的热狗折叠酰基辅酶 A 硫酯酶结构域和一个未知折叠和功能的 N 端结构域组成。hTHEM4 与 Akt1 调节和细胞凋亡有关。在此，我们报告了与人 THEM4 结合的十一烷-2-酮-CoA 的 X 射线结构。结构引导的突变分析用于确认催化残基。N 端结构域部分由不规则和灵活的二级结构组成，类似于蛋白质结合结构域。我们通过免疫沉淀和 Akt1 激酶活性的抑制来证明 hTHEM4 与 Akt1 的直接结合，从而提供了独立的证据表明 hTHEM4 是 Akt1 的负调节剂。
