Identification and characterization of the a-CA in the outer membrane vesicles produced by Helicobacter pylori

The genome of Helicobacter pylori encodes for carbonic anhydrases (CAs, EC 4.2.1.1) belonging to the aand b-CA classes, which together with urease, have a pivotal role in the acid acclimation of the microorganism within the human stomach. Recently, in the exoproteome of H. pylori, a CA with no indication of the corresponding class was identified. Here, using the protonography and the mass spectrometry, a CA belonging to the a-class was detected in the outer membrane vesicles (OMVs) generated by planktonic and biofilm phenotypes of four H. pylori strains. The amount of this metalloenzyme was higher in the planktonic OMVs (pOMVs) than in the biofilm OMVs (bOMVs). Furthermore, the content of a-CA increases over time in the pOMVs. The identification of the a-CA in pOMVs and bOMVs might shed new light on the role of this enzyme in the colonization, survival, persistence, and pathogenesis of H. pylor

The Evolution of Pepsinogen C Genes in Vertebrates: Duplication, Loss and Functional Diversification

<div><h3>Background</h3><p>Aspartic proteases comprise a large group of enzymes involved in peptide proteolysis. This collection includes prominent enzymes globally categorized as pepsins, which are derived from pepsinogen precursors. Pepsins are involved in gastric digestion, a hallmark of vertebrate physiology. An important member among the pepsinogens is pepsinogen C (<em>Pgc</em>). A particular aspect of <em>Pgc</em> is its apparent single copy status, which contrasts with the numerous gene copies found for example in pepsinogen A (<em>Pga</em>). Although gene sequences with similarity to <em>Pgc</em> have been described in some vertebrate groups, no exhaustive evolutionary framework has been considered so far.</p> <h3>Methodology/Principal Findings</h3><p>By combining phylogenetics and genomic analysis, we find an unexpected <em>Pgc</em> diversity in the vertebrate sub-phylum. We were able to reconstruct gene duplication timings relative to the divergence of major vertebrate clades. Before tetrapod divergence, a single <em>Pgc</em> gene tandemly expanded to produce two gene lineages (<em>Pgbc</em> and <em>Pgc2</em>). These have been differentially retained in various classes. Accordingly, we find <em>Pgc2</em> in sauropsids, amphibians and marsupials, but not in eutherian mammals. <em>Pgbc</em> was retained in amphibians, but duplicated in the ancestor of amniotes giving rise to <em>Pgb</em> and <em>Pgc1</em>. The latter was retained in mammals and probably in reptiles and marsupials but not in birds. <em>Pgb</em> was kept in all of the amniote clade with independent episodes of loss in some mammalian species. Lineage specific expansions of <em>Pgc2</em> and <em>Pgbc</em> have also occurred in marsupials and amphibians respectively. We find that teleost and tetrapod <em>Pgc</em> genes reside in distinct genomic regions hinting at a possible translocation.</p> <h3>Conclusions</h3><p>We conclude that the repertoire of <em>Pgc</em> genes is larger than previously reported, and that tandem duplications have modelled the history of <em>Pgc</em> genes. We hypothesize that gene expansion lead to functional divergence in tetrapods, coincident with the invasion of terrestrial habitats.</p> </div