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.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Acyl Coenzyme A / chemistry
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Acyl Coenzyme A / metabolism
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Adaptor Proteins, Signal Transducing / chemistry*
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Adaptor Proteins, Signal Transducing / metabolism
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Catalytic Domain / drug effects
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Crystallography, X-Ray
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Humans
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Membrane Proteins / chemistry*
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Membrane Proteins / metabolism
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Models, Molecular
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Protein Structure, Secondary
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Proto-Oncogene Proteins c-akt / chemistry
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Proto-Oncogene Proteins c-akt / metabolism
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Thiolester Hydrolases / antagonists & inhibitors
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Thiolester Hydrolases / chemistry*
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Thiolester Hydrolases / metabolism
Substances
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Acyl Coenzyme A
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Adaptor Proteins, Signal Transducing
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Membrane Proteins
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Proto-Oncogene Proteins c-akt
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THEM4 protein, human
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Thiolester Hydrolases