title: C-terminal end and aminoacid Lys48 in HMG-CoA lyase are involved in substrate binding and enzyme activity.
abstract: 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) lyase adopts a (betaalpha)(8) TIM barrel structure with an additional beta9, alpha11 and alpha12 helices. Location of HMG part of the substrate has been suggested but the binding mode for the CoA moiety remains to be resolved. As mutation F305 fs(-2), which involves the last 21 residues of the protein, and mutation K48N caused 3-hydroxy-3-methylglutaric aciduria in two patients, we examined the role of the C-terminal end and Lys(48) in enzyme activity. Expression studies of various C-terminal-end-deleted and K48N-mutated proteins revealed that residues 311-313 (localized in the loop between alpha11 and alpha12 helices) and Lys(48) are essential for enzyme activity. An in silico docking model locating HMG-CoA on the surface of the enzyme implicates Asn(311) and Lys(313) in substrate binding by establishing multiple polar contacts with phosphate and ribose groups of adenosine, and Lys(48) by contacting the carboxyl group of the panthotenic acid moiety.
authors: Department of Biochemistry and Molecular Biology, School of Health Sciences, Universitat Internacional de Catalunya, E-08195 Sant Cugat, Barcelona, Spain.
authors: Patricia
authors: P
authors: Carrasco
authors: Sebastian
authors: S
authors: Menao
authors: Eduardo
authors: E
authors: López-Viñas
authors: Gabriel
authors: G
authors: Santpere
authors: Josep
authors: J
authors: Clotet
authors: Adriana Y
authors: AY
authors: Sierra
authors: Esther
authors: E
authors: Gratacós
authors: Beatriz
authors: B
authors: Puisac
authors: Paulino
authors: P
authors: Gómez-Puertas
authors: Fausto G
authors: FG
authors: Hegardt
authors: Juan
authors: J
authors: Pie
authors: Núria
authors: N
authors: Casals
mesh_terms: D000214:Acyl Coenzyme A / Q000737:chemistry*; D000595:Amino Acid Sequence; D017124:Conserved Sequence; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D007652:Oxo-Acid-Lyases / Q000096:biosynthesis / Q000737:chemistry* / Q000172:deficiency / Q000235:genetics*; D011485:Protein Binding; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics
chemical_list: D000214:Acyl Coenzyme A; D011994:Recombinant Proteins; C008047:3-hydroxy-3-methylglutaryl-coenzyme A; D007652:Oxo-Acid-Lyases; C021410:3-hydroxy-3-methylglutaryl-coenzyme A lyase