title: Molecular cloning of CoA Synthase. The missing link in CoA biosynthesis.
abstract: Coenzyme A functions as a carrier of acetyl and acyl groups in living cells and is essential for numerous biosynthetic, energy-yielding, and degradative metabolic pathways. There are five enzymatic steps in CoA biosynthesis. To date, molecular cloning of enzymes involved in the CoA biosynthetic pathway in mammals has been only reported for pantothenate kinase. In this study, we present cDNA cloning and functional characterization of CoA synthase. It has an open reading frame of 563 aa and encodes a protein of approximately 60 kDa. Sequence alignments suggested that the protein possesses both phosphopantetheine adenylyltransferase and dephospho-CoA kinase domains. Biochemical assays using wild type recombinant protein confirmed the gene product indeed contained both these enzymatic activities. The presence of intrinsic phosphopantetheine adenylyltransferase activity was further confirmed by site-directed mutagenesis. Therefore, this study describes the first cloning and characterization of a mammalian CoA synthase and confirms this is a bifunctional enzyme containing the last two components of CoA biosynthesis.
authors: Department of Structure and Function of Nucleic Acid, The Institute of Molecular Biology and Genetics, 150 Zabolotnogo Street, Kyiv 143, Ukraine.
authors: Alexander
authors: A
authors: Zhyvoloup
authors: Ivan
authors: I
authors: Nemazanyy
authors: Aleksei
authors: A
authors: Babich
authors: Ganna
authors: G
authors: Panasyuk
authors: Natalya
authors: N
authors: Pobigailo
authors: Mariya
authors: M
authors: Vudmaska
authors: Valeriy
authors: V
authors: Naidenov
authors: Oleksandr
authors: O
authors: Kukharenko
authors: Sergiy
authors: S
authors: Palchevskii
authors: Liliya
authors: L
authors: Savinska
authors: Galina
authors: G
authors: Ovcharenko
authors: Frederique
authors: F
authors: Verdier
authors: Taras
authors: T
authors: Valovka
authors: Tim
authors: T
authors: Fenton
authors: Heike
authors: H
authors: Rebholz
authors: Mong-Lien
authors: ML
authors: Wang
authors: Peter
authors: P
authors: Shepherd
authors: Genadiy
authors: G
authors: Matsuka
authors: Valeriy
authors: V
authors: Filonenko
authors: Ivan T
authors: IT
authors: Gout
mesh_terms: D000106:Acetate-CoA Ligase / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D002460:Cell Line; D003001:Cloning, Molecular; D003065:Coenzyme A / Q000096:biosynthesis*; D018076:DNA, Complementary / Q000378:metabolism; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids / Q000378:metabolism; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014018:Tissue Distribution; D014166:Transferases / Q000235:genetics*; D020798:Two-Hybrid System Techniques
chemical_list: D018076:DNA, Complementary; D014166:Transferases; C509687:COASY protein, human; D000106:Acetate-CoA Ligase; D003065:Coenzyme A