Literature summary for 6.2.1.16 extracted from

Hasegawa, S.; Inoue, D.; Yamasaki, M.; Li, C.; Imai, M.; Takahashi, N.; Fukui, T.
Site-specific cleavage of acetoacetyl-CoA synthetase by legumain (2016), FEBS Lett., 590, 1592-1601 .

Search for more literature for 6.2.1.16

Cloned(Commentary)

Cloned (Comment)	Organism
gene Aacs, recombinant expression of FLAG-tagged enzyme in Lenti-X-293T cells, overexpression of wild-type and mutant enzymes in HEK-293 cells	Mus musculus

Protein Variants

Protein Variants	Comment	Organism
N320Q	site-directed mutagenesis, the mutant is not cleaved by legumain	Mus musculus
N449Q	site-directed mutagenesis, the mutant is not cleaved by legumain	Mus musculus
N500Q	site-directed mutagenesis, the mutant is cleaved by legumain	Mus musculus
N503Q	site-directed mutagenesis, the mutant iscleaved by legumain, but the mutation abolishes the formation of the 56-kDa band through cleavage by legumain	Mus musculus
N545Q	site-directed mutagenesis, catalytically inactive mutant, that is cleaved by legumain	Mus musculus
N547Q	site-directed mutagenesis, the mutant iscleaved by legumain, but the mutation abolishes the formation of the 56-kDa band through cleavage by legumain	Mus musculus

General Stability

General Stability	Organism
acetoacetyl-CoA synthetase (AACS) is cleaved by legumain, a lysosomal asparaginyl endopeptidase. Asn547 is the specific cleavage site of AACS in mouse livers. The cleaved form of AACS (1-547) loses the ability to convert acetoacetate to acetoacetyl-CoA	Mus musculus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + acetoacetate + CoA	Mus musculus	-	AMP + diphosphate + acetoacetyl-CoA	-	?

Organism

Organism	UniProt	Comment	Textmining
Mus musculus	Q9D2R0	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
proteolytic modification	site-specific cleavage at residue AACS Asn503 and Asn547 of acetoacetyl-CoA synthetase by recombinant autoactivated legumain, a lysosomal asparaginyl endopeptidase, at pH 5.0 and 30°C. The cleaved form of AACS (1-547) loses the ability to convert acetoacetate to acetoacetyl-CoA. AACS is cleaved by legumain in the liver and the kidney to give two primary bands of approximately 56 kDa and 48 kDa, AACS (1-547) and AACS (1-503) are approximately 56 kDa and 55 kDa, respectively. The cleavage site for the formation of the 56-kDa product is located in the C-terminal side region from amino acid 487, whereas that of the 48-kDa product is located in the N-terminal side region from amino acid 468	Mus musculus

Purification (Commentary)

Purification (Comment)	Organism
-	Mus musculus
recombinant FLAG-tagged enzyme from Lenti-X-293T cells by affinity chromatography and ultrafiltration	Mus musculus

Source Tissue

Source Tissue	Comment	Organism	Textmining
commercial preparation	-	Mus musculus	-
kidney	-	Mus musculus	-
liver	-	Mus musculus	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + acetoacetate + CoA	-	Mus musculus	AMP + diphosphate + acetoacetyl-CoA	-	?

Synonyms

Synonyms	Comment	Organism
AACS	-	Mus musculus
Acetoacetyl-CoA synthetase	-	Mus musculus

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Mus musculus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	assay at	Mus musculus

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Mus musculus

General Information

General Information	Comment	Organism
malfunction	overexpression of recombinant mutants N500Q, N503Q, or N547Q, as well as of the wild-type enzyme, increases the ketone body-utilizing activity of HEK-293 cells, but that of N545Q does not. Overexpression of wild-type AACS, N500Q, or N503Q has no effect on legumain activity, but mutations N545Q and N547Q significantly reduce the activity compared to wild-type	Mus musculus
metabolism	acetoacetyl-CoA synthetase (AACS) is responsible for the synthesis of cholesterol and fatty acids. It is cleaved by legumain, a lysosomal asparaginyl endopeptidase. Asn547 is the specific cleavage site of AACS in mouse livers. The cleaved form of AACS (1-547) loses the ability to convert acetoacetate to acetoacetyl-CoA. Overexpression of the cleaved form of AACS (1-547) increases the protein expression of caveolin-1, the principal component of the caveolae. Cleavage of AACS by legumain is critical for the regulation of enzymatic activity and results in gain-of-function changes	Mus musculus
additional information	site-specific cleavage at residue Asn547 of acetoacetyl-CoA synthetase by legumain, a lysosomal asparaginyl endopeptidase. The cleaved form of AACS (1-547) loses the ability to convert acetoacetate to acetoacetyl-CoA	Mus musculus
physiological function	acetoacetyl-CoA synthetase (AACS) is a ketone body-utilizing enzyme and is responsible for the synthesis of cholesterol and fatty acids. Overexpression of wild-type AACS and AACS (1-547) increased the protein expression of caveolin-1, a scaffolding protein and the principal component of the caveolae, in the cytosol of liver cells. Enzyme AACS has a unique role in caveolae/lipid rafts	Mus musculus
physiological function	hydrodynamics-based gene transduction shows that overexpression of AACS (1547) increases the protein expression of caveolin-1, the principal component of the caveolae. Cleavage of AACS by legumain is critical for the regulation of enzymatic activity and results in gain-of-function changes	Mus musculus

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Release 2023.1 (January 2023)