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PMID: 9401030 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Characterization of nicotinamide mononucleotide adenylyltransferase from thermophilic archaea.

Journal of bacteriology ·Vol. 179 ·No. 24 ·1997-12-00 ·Pages 7718-23

Raffaelli N, Pisani FM, Lorenzi T, Emanuelli M, Amici A, Ruggieri S, Magni G

Abstract

The enzyme nicotinamide mononucleotide (NMN) adenylyltransferase (EC 2.7.7.1) catalyzes the synthesis of NAD+ and nicotinic acid adenine dinucleotide. It has been purified to homogeneity from cellular extracts of the thermophilic archaeon Sulfolobus solfataricus. Through a database search, a highly significant match was found between its N-terminal sequence and a hypothetical protein coded by the thermophilic archaeon Methanococcus jannaschii MJ0541 open reading frame (GenBank accession no. U67503). The MJ0541 gene was isolated, cloned into a T7-based vector, and expressed in Escherichia coli cells, yielding a high level of thermophilic NMN adenylyltransferase activity. The expressed protein was purified to homogeneity by a single-step chromatographic procedure. Both the subunit molecular mass and the N-terminal sequence of the pure recombinant protein were as expected from the deduced amino acid sequence of the MJ0541 open reading frame-encoded protein. Molecular and kinetic properties of the enzymes from both archaea are reported and compared with those already known for the mesophilic eukaryotic NMN adenylyltransferase.

MeSH Terms
Amino Acid Sequence Amino Acids/analysis Archaeal Proteins/genetics,isolation & purification Cloning, Molecular Enzyme Stability Escherichia coli/genetics Hot Temperature Methanococcus/enzymology,genetics Molecular Sequence Data Nicotinamide-Nucleotide Adenylyltransferase/genetics,isolation & purification Recombinant Proteins/isolation & purification Sequence Analysis Sequence Homology, Amino Acid Sulfolobus/enzymology
Chemicals
Amino Acids Archaeal Proteins Recombinant Proteins Nicotinamide-Nucleotide Adenylyltransferase
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Raffaelli N
Istituto di Biochimica, Facoltà di Medicina e Chirurgia, Università di Ancona, Italy.
Pisani F M
Lorenzi T
Emanuelli M
Amici A
Ruggieri S
Magni G
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.179.24.7718-7723.1997
Published
1997-12-00
Pages
7718-23
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC179734
Subset
IM
Databases
GENBANK
U67503
External Links
PubMed source DOI: 10.1128/jb.179.24.7718-7723.1997 PMC Full Text View journal details
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