Involvement of Chromosomally Encoded Homologs of the RRNPP Protein Family in Enterococcus faecalis Biofilm Formation and Urinary Tract Infection Pathogenesis.

is an opportunistic pathogen capable of causing infections, including endocarditis and urinary tract infections (UTI). One of the well-characterized quorum-sensing pathways in involves coordination of the conjugal transfer of pheromone-responsive plasmids by PrgX, a member of the RRNPP protein family. Members of this protein family in various have also been shown to contribute to numerous cellular processes, including sporulation, competence, conjugation, nutrient sensing, biofilm formation, and virulence.

Confined Mobility of TonB and FepA in Escherichia coli Membranes.

The important process of nutrient uptake in Escherichia coli, in many cases, involves transit of the nutrient through a class of beta-barrel proteins in the outer membrane known as TonB-dependent transporters (TBDTs) and requires interaction with the inner membrane protein TonB. Here we have imaged the mobility of the ferric enterobactin transporter FepA and TonB by tracking them in the membranes of live E. coli with single-molecule resolution at time-scales ranging from milliseconds to seconds.

High-Throughput Screening Assay for Inhibitors of TonB-Dependent Iron Transport.

The TonB-dependent Gram-negative bacterial outer membrane protein FepA actively transports the siderophore ferric enterobactin (FeEnt) into the periplasm. We developed a high-throughput screening (HTS) assay that observes FeEnt uptake through FepA in living Escherichia coli, by monitoring fluorescence quenching that occurs upon binding of FeEnt, and then unquenching as the bacteria deplete it from solution by transport. We optimized the labeling and spectroscopic methods to screen for inhibitors of TonB-dependent iron uptake through the outer membrane.

Concerted loop motion triggers induced fit of FepA to ferric enterobactin.

Spectroscopic analyses of fluorophore-labeled Escherichia coli FepA described dynamic actions of its surface loops during binding and transport of ferric enterobactin (FeEnt). When FeEnt bound to fluoresceinated FepA, in living cells or outer membrane fragments, quenching of fluorophore emissions reflected conformational motion of the external vestibular loops. We reacted Cys sulfhydryls in seven surface loops (L2, L3, L4, L5, L7 L8, and L11) with fluorophore maleimides.

Energy-dependent motion of TonB in the Gram-negative bacterial inner membrane.

Gram-negative bacteria acquire iron with TonB-dependent uptake systems. The TonB-ExbBD inner membrane complex is hypothesized to transfer energy to outer membrane (OM) iron transporters. Fluorescence microscopic characterization of green fluorescent protein (GFP)-TonB hybrid proteins revealed an unexpected, restricted localization of TonB in the cell envelope.

Role of catecholate siderophores in gram-negative bacterial colonization of the mouse gut.

We investigated the importance of the production of catecholate siderophores, and the utilization of their iron (III) complexes, to colonization of the mouse intestinal tract by Escherichia coli. First, a ΔtonB strain was completely unable to colonize mice. Next, we compared wild type E.

Evidence of herpesvirus infection in Woodland Caribou in Saskatchewan.

Sera were collected from 40 female and two male woodland caribou (Rangifer tarandus caribou) in Saskatchewan (Canada) from March 1992 to January 1995, inclusive. The samples were examined for antibodies against smooth Brucella spp., five serovars of Leptospira interrogans, bovine viral diarrhea virus, and bovine herpesvirus 1 (BHV-1).

A blocking ELISA with improved sensitivity for the detection of passively acquired maternal antibodies to BHV-1.

Four serological tests were evaluated for their ability to detect passively acquired maternal antibodies to Bovine herpesvirus 1. A blocking enzyme-linked immunosorbent assay demonstrated superior sensitivity in the detection of such antibodies in calves up to 9-11 months old, versus calves up to 7 months old for other tests.