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Characterization of ATPase activity of the AAA ARC from Bifidobacterium longum subsp. infantis

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Guzmán-Rodríguez M., de la Rosa A., Santos L.

2015

vol. 62

issue 2

221-227

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.

HLA-Cw and TCR vβ analysis in twenty Mexican patients with psoriasis

64%

Santos L., León-Galvan M., Fuentes-Ahumada C., Marino-Marmolejo E., León-Rodríguez A., Rosa A., Ceballos-Salazar R., Torres-Alvarez B., Moncada-González B.

Open Medicine

2011

vol. 6

issue 4

442-448

Genetic background and T-cell expansion have been confirmed as the most important factors leading to psoriasis susceptibility in the Caucasian population. This study was performed to identify the T-cell receptor Vβ repertoire and HLA-Cw genotype in twenty Mexicans of two different ethnicities with severe chronic plaque-type psoriasis. HLA-Cw typing was performed to detect the allele pattern by SSP-PCR. In parallel, RT-PCR and Western blot were used for the identification of the TCR Vβ expression in peripheral blood cells. We identified a variety of HLA-Cw alleles in this group of patients distinct from the widely known HLA-Cw 0602 Caucasian allele. Moreover, TCR Vβ-2 and Vβ-7 clone-type frequencies were different and statistically significant (P = 0.0280). We speculate that because of diverse genetic backgrounds, the susceptibility to disease and activation of T-cells for a proper immune response could be specific; therefore, the findings might contribute to the elucidation of the pathogenesis in psoriatic Mexican patients.

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1 Acta Biochimica Polonica

1 Open Medicine

1 2015

1 2011

2 Santos L.

1 Ceballos-Salazar R.

1 Fuentes-Ahumada C.

1 Guzmán-Rodríguez M.

1 León-Galvan M.

1 León-Rodríguez A.

1 Marino-Marmolejo E.

1 Moncada-González B.

1 Rosa A.

1 Torres-Alvarez B.

1 de la Rosa A.

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Characterization of ATPase activity of the AAA ARC from Bifidobacterium longum subsp. infantis

HLA-Cw and TCR vβ analysis in twenty Mexican patients with psoriasis