Synechocystis sp. slr0787 protein is a novel bifunctional enzyme endowed with both nicotinamide mononucleotide adenylyltransferase and 'Nudix' hydrolase activities.
Synechocystis sp. slr0787 open reading frame encodes a 339 residue polypeptide with a predicted molecular mass of 38.5 kDa. Its deduced amino acid sequence shows extensive homology with known separate sequences of proteins from the thermophilic archaeon Methanococcus jannaschii. The N-terminal domain is highly homologous to the archaeal NMN adenylyltransferase, which catalyzes NAD synthesis from NMN and ATP. The C-terminal domain shares homology with the archaeal ADP-ribose pyrophosphatase, a member of the 'Nudix' hydrolase family. The slr0787 gene has been cloned into a T7-based vector for expression in Escherichia coli cells. The recombinant protein has been purified to homogeneity and demonstrated to possess both NMN adenylyltransferase and ADP-ribose pyrophosphatase activities. Both activities have been characterized and compared to their archaeal counterparts.
Raffaelli
N
N
Istituto di Biochimica, Facoltà di Medicina e Chirurgia, University of Ancona, Italy.
Lorenzi
T
T
Amici
A
A
Emanuelli
M
M
Ruggieri
S
S
Magni
G
G
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D003001:Cloning, Molecular; D000458:Cyanobacteria / Q000201:enzymology*; D004795:Enzyme Stability; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D017017:Methanococcus / Q000201:enzymology; D008969:Molecular Sequence Data; D009612:Nicotinamide-Nucleotide Adenylyltransferase / Q000737:chemistry / Q000235:genetics*; D011755:Pyrophosphatases / Q000737:chemistry / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013696:Temperature
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D008345:Manganese; D009612:Nicotinamide-Nucleotide Adenylyltransferase; C117989:SLR0787 protein, Synechocystis; D011755:Pyrophosphatases; D008274:Magnesium