title: Site-specific cleavage of acetoacetyl-CoA synthetase by legumain.
abstract: Acetoacetyl-CoA synthetase (AACS) is a ketone body-utilizing enzyme and is responsible for the synthesis of cholesterol and fatty acids. We have previously shown that AACS is cleaved by legumain, a lysosomal asparaginyl endopeptidase. In this study, we attempted to determine the cleavage site of AACS. Mutagenesis analysis of AACS revealed that Asn547 is the specific cleavage site of AACS in mouse livers. The cleaved form of AACS (1-547) lost the ability to convert acetoacetate to acetoacetyl-CoA. Moreover, hydrodynamics-based gene transduction showed that overexpression of AACS (1-547) increases the protein expression of caveolin-1, the principal component of the caveolae. These results suggest that cleavage of AACS by legumain is critical for the regulation of enzymatic activity and results in gain-of-function changes.
authors.affiliation: Department of Health Chemistry, Hoshi University, Shinagawa, Tokyo, Japan.
authors.forename: Shinya
authors.initials: S
authors.lastname: Hasegawa
authors.affiliation: Department of Health Chemistry, Hoshi University, Shinagawa, Tokyo, Japan.
authors.forename: Daiki
authors.initials: D
authors.lastname: Inoue
authors.affiliation: Department of Health Chemistry, Hoshi University, Shinagawa, Tokyo, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Yamasaki
authors.affiliation: Laboratory of Physiological Chemistry, Institute of Medicinal Chemistry, Hoshi University, Shinagawa, Tokyo, Japan.
authors.forename: Chuan
authors.initials: C
authors.lastname: Li
authors.affiliation: Laboratory of Physiological Chemistry, Institute of Medicinal Chemistry, Hoshi University, Shinagawa, Tokyo, Japan.
authors.forename: Masahiko
authors.initials: M
authors.lastname: Imai
authors.affiliation: Laboratory of Physiological Chemistry, Institute of Medicinal Chemistry, Hoshi University, Shinagawa, Tokyo, Japan.
authors.forename: Noriko
authors.initials: N
authors.lastname: Takahashi
authors.affiliation: Department of Pharmacy, College of Pharmaceutical Sciences, Ritsumeikan University, Kyoto, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Fukui
mesh_terms: D000818:Animals; D001665:Binding Sites; D002478:Cells, Cultured; D003066:Coenzyme A Ligases / Q000737:chemistry* / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D004789:Enzyme Activation; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D054730:Protein Interaction Domains and Motifs; D059748:Proteolysis; D013379:Substrate Specificity; D000099240:Asparaginyl Endopeptidase
chemical_list: D003066:Coenzyme A Ligases; D003546:Cysteine Endopeptidases; D000099240:Asparaginyl Endopeptidase; C023747:acetoacetyl-CoA synthetase
keywords: acetoacetyl-CoA synthetase; caveolin-1; ketone body; legumain; mutagenesis