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Type II fatty acid biosynthesis systems are essential for membrane formation in bacteria, making the constituent proteins of this pathway attractive targets for antibacterial drug discovery. The third step in the elongation cycle of the type II fatty acid biosynthesis is catalyzed by ␤-hydroxyacyl-(acyl carrier protein) (ACP) dehydratase. There are two isoforms. FabZ, which catalyzes the dehydration of (3R)-hydroxyacyl-ACP to trans-2-acyl-ACP, is a universally expressed component of the bacterial type II system. FabA, the second isoform, as has more limited distribution in nature and, in addition to dehydration, also carries out the isomerization of trans-2-to cis-3-decenoyl-ACP as an essential step in unsaturated fatty acid biosynthesis. We report the structure of FabZ from the important human pathogen Pseudomonas aeruginosa at 2.5 Å of resolution. PaFabZ is a hexamer (trimer of dimers) with the His/Glu catalytic dyad located within a deep, narrow tunnel formed at the dimer interface. Site-directed mutagenesis experiments showed that the obvious differences in the active site residues that distinguish the FabA and FabZ subfamilies of dehydratases do not account for the unique ability of FabA to catalyze isomerization. Because the catalytic machinery of the two enzymes is practically indistinguishable, the structural differences observed in the shape of the substrate binding channels of FabA and FabZ lead us to hypothesize that the different shapes of the tunnels control the conformation and positioning of the bound substrate, allowing FabA, but not FabZ, to catalyze the isomerization reaction.In eubacteria and their endosymbiotic descendants (the plastids of plants and apicomplexan parasites) fatty acids are produced by what is known as the type II fatty acid biosynthetic pathway (1-3). The steps in this pathway are catalyzed by a universal set of enzymes, each encoded by a separate gene, that have been most closely studied in the model organism Escherichia coli (4, 5). The growing acyl chain is shuttled between the pathway enzymes attached to the 4Ј-phosphopantetheine prosthetic group of a dedicated carrier protein, ACP.1 This system contrasts with the type I fatty acid biosynthesis system that exists in metazoans, where multifunctional polypeptide chains (6) encode all activities in chain initiation and elongation. The intermediates in the type I system are shuffled from one catalytic site to another without being released from the complex. In light of the profound differences in these two systems, enzymes of the type II pathway have emerged as attractive targets for the development of novel antimicrobial or antiparasitic agents (2,7,8). The core feature of the type II pathway is the fatty acid elongation cycle, which extends the fatty acid chain by two carbons in each round. There are four steps in the cycle, and the proteins involved in E. coli are 1) condensation of malonyl-ACP with acyl-ACP, catalyzed by the FabB-and FabF-condensing enzymes, 2) reduction of the ␤-keto moiety by the NADPH-dependent FabG re...

This article provides a review of the contribution of Axel Honneth's model of recognition for critical social work. While Honneth's tripartite conceptualisation of optimal identityformation is positively appraised, his analysis of the link between misrecognition, the experience of shame and eventual sense of moral outrage, is contested. Drawing on a range of sources, including the sociology of shame, Honneth's ideas about the emotional antecedents of emancipatory action are revised to guide critical social work with misrecognised service users.

Treponema pallidum, the causative agent of syphilis, is a highly invasive pathogenic spirochete capable of attaching to host cells, invading the tissue barrier, and undergoing rapid widespread dissemination via the circulatory system. The T. pallidum adhesin Tp0751 was previously shown to bind laminin, the most abundant component of the basement membrane, suggesting a role for this adhesin in host tissue colonization and bacterial dissemination. We hypothesized that similar to that of other invasive pathogens, the interaction of T. pallidum with host coagulation proteins, such as fibrinogen, may also be crucial for dissemination via the circulatory system. To test this prediction, we used enzyme-linked immunosorbent assay (ELISA) methodology to demonstrate specific binding of soluble recombinant Tp0751 to human fibrinogen. Click-chemistry-based palmitoylation profiling of heterologously expressed Tp0751 confirmed the presence of a lipid attachment site within this adhesin. Analysis of the Tp0751 primary sequence revealed the presence of a C-terminal putative HEXXH metalloprotease motif, and in vitro degradation assays confirmed that recombinant Tp0751 purified from both insect and Escherichia coli expression systems degrades human fibrinogen and laminin. The proteolytic activity of Tp0751 was abolished by the presence of the metalloprotease inhibitor 1,10-phenanthroline. Further, inductively coupled plasma-mass spectrometry showed that Tp0751 binds zinc and calcium. Collectively, these results indicate that Tp0751 is a zinc-dependent, membrane-associated protease that exhibits metalloprotease-like characteristics. However, site-directed mutagenesis of the HEXXH motif to HQXXH did not abolish the proteolytic activity of Tp0751, indicating that further mutagenesis studies are required to elucidate the critical active site residues associated with this protein. This study represents the first published description of a T. pallidum protease capable of degrading host components and thus provides novel insight into the mechanism of T. pallidum dissemination.