title: Degradation of acetoacetyl-CoA synthetase, a ketone body-utilizing enzyme, by legumain in the mouse kidney.
abstract: Acetoacetyl-CoA synthetase (AACS) is a ketone body-utilizing enzyme, which is responsible for the synthesis of cholesterol and fatty acids from ketone bodies in lipogenic tissues, such as the liver and adipocytes. To explore the possibility of AACS regulation at the protein-processing level, we investigated the proteolytic degradation of AACS. Western blot analysis showed that the 75.1kDa AACS was cleaved to form a protein of approximately 55kDa in the kidney, which has considerable high activity of legumain, a lysosomal asparaginyl endopeptidase. Co-expression of AACS and legumain in HEK 293 cells generated the 55kDa product from AACS. Moreover, incubation of recombinant AACS with recombinant legumain resulted in the degradation of AACS. Knockdown of legumain with short-hairpin RNA against legumain using the hydrodynamics method led to a decrease in the 55kDa band of AACS in mouse kidney. These results suggest that legumain is involved in the processing of AACS through the lysosomal degradation pathway in the kidney.
authors.affiliation: Department of Health Chemistry, Hoshi University, Ebara, Shinagawa, Tokyo 142-8501, Japan. Electronic address: s-hasegawa@hoshi.ac.jp.
authors.forename: Shinya
authors.initials: S
authors.lastname: Hasegawa
authors.affiliation: Department of Health Chemistry, Hoshi University, Ebara, Shinagawa, Tokyo 142-8501, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Yamasaki
authors.affiliation: Department of Health Chemistry, Hoshi University, Ebara, Shinagawa, Tokyo 142-8501, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Fukui
mesh_terms: D000818:Animals; D001483:Base Sequence; D003066:Coenzyme A Ligases / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000494:pharmacology*; D017931:DNA Primers; D057809:HEK293 Cells; D006801:Humans; D007657:Ketone Bodies / Q000378:metabolism*; D007668:Kidney / Q000187:drug effects*; D008297:Male; D051379:Mice; D059748:Proteolysis; D000099240:Asparaginyl Endopeptidase
chemical_list: D003066:Coenzyme A Ligases; D003546:Cysteine Endopeptidases; D017931:DNA Primers; D007657:Ketone Bodies; D000099240:Asparaginyl Endopeptidase; C023747:acetoacetyl-CoA synthetase
keywords: Acetoacetyl-CoA synthetase; Ketone body; Kidney; Legumain; Proteolytic degradation