title: Characterization of ATPase activity of the AAA ARC from Bifidobacterium longum subsp. infantis.
abstract: Bifidobacteria are considered to be probiotics that exist in the large intestine and are helpful to maintain human health. Oral administration of bifidobacteria may be effective in improving the intestinal flora and environment, stimulating the immune response and possibly preventing cancer. However, for consistent and positive results, further well-controlled studies are urgently needed to describe the basic mechanisms of this microorganism. Analysis of the proteasome-lacking Bifidobacterium longum genome reveals that it possesses a gene, IPR003593 AAA ATPase core, which codes a 56 kDa protein containing one AAA ATPase domain. Phylogenetic classification made by CLANS, positioned this sequence into the ARC divergent branch of the AAA ATPase family of proteins. N-terminal analysis of the sequence indicates this protein is closely related to other ATPases such as the Rhodococcus erythropolis ARC, Archaeoglobus fulgidus PAN, Mycobacterium tuberculosis Mpa and the human proteasomal Rpt1 subunit. This gene was cloned, the full-length recombinant protein was overexpressed in Escherichia coli, purified as a high-molecular size complex and named Bl-ARC. Enzymatic characterization showed that Bl-ARC ATPase is active, Mg(+2)-dependent and sensitive to N-ethylmaleimide. Gene organization positions bl-arc in a region flanked by a cluster of genes that includes pup, dop and pafA genes. These findings point to a possible function as a chaperone in the degradation pathway via pupylation.
authors.affiliation: División de Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica, A.C. (IPICYT), CP 78216 San Luis Potosí, México.
authors.forename: Mabel
authors.initials: M
authors.lastname: Guzmán-Rodríguez
authors.affiliation: División de Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica, A.C. (IPICYT), CP 78216 San Luis Potosí, México.
authors.forename: Ana Paulina Barba
authors.initials: AP
authors.lastname: de la Rosa
authors.affiliation: División de Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica, A.C. (IPICYT), CP 78216 San Luis Potosí, México.
authors.forename: Leticia
authors.initials: L
authors.lastname: Santos
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001644:Bifidobacterium / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D005033:Ethylmaleimide / Q000494:pharmacology; D006868:Hydrolysis; D008274:Magnesium / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D000251:Adenosine Triphosphatases; D008274:Magnesium; D005033:Ethylmaleimide