title: Insights into ligand binding and catalysis of a central step in NAD+ synthesis: structures of Methanobacterium thermoautotrophicum NMN adenylyltransferase complexes.
abstract: Nicotinamide mononucleotide adenylyltransferase (NMNATase) catalyzes the linking of NMN(+) or NaMN(+) with ATP, which in all organisms is one of the common step in the synthesis of the ubiquitous coenzyme NAD(+), via both de novo and salvage biosynthetic pathways. The structure of Methanobacterium thermoautotrophicum NMNATase determined using multiwavelength anomalous dispersion phasing revealed a nucleotide-binding fold common to nucleotidyltransferase proteins. An NAD(+) molecule and a sulfate ion were bound in the active site allowing the identification of residues involved in product binding. In addition, the role of the conserved (16)HXGH(19) active site motif in catalysis was probed by mutagenic, enzymatic and crystallographic techniques, including the characterization of an NMN(+)/SO4(2-) complex of mutant H19A NMNATase.
authors.affiliation: Division of Molecular and Structural Biology, Ontario Cancer Institute, 610 University Avenue Toronto M5G 2M9, Ontario, Canada.
authors.forename: V
authors.initials: V
authors.lastname: Saridakis
authors.forename: D
authors.initials: D
authors.lastname: Christendat
authors.forename: M S
authors.initials: MS
authors.lastname: Kimber
authors.forename: A
authors.initials: A
authors.lastname: Dharamsi
authors.forename: A M
authors.initials: AM
authors.lastname: Edwards
authors.forename: E F
authors.initials: EF
authors.lastname: Pai
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002384:Catalysis; D020134:Catalytic Domain; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D008024:Ligands; D017014:Methanobacterium / Q000201:enzymology*; D008956:Models, Chemical; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D009243:NAD / Q000096:biosynthesis*; D009612:Nicotinamide-Nucleotide Adenylyltransferase / Q000737:chemistry* / Q000235:genetics; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017386:Sequence Homology, Amino Acid
chemical_list: D008024:Ligands; D009243:NAD; D009612:Nicotinamide-Nucleotide Adenylyltransferase