The flagellum of Methanococcus voltae is composed four structural flagellin proteins FlaA, FlaB1, FlaB2, and FlaB3. These possess a total 15 potential N-linked sequons (NX(S/T)) show mass shift on an SDS-polyacrylamide gel indicating significant post-translational modification. We describe here the characterization glycan from M. using spectrometry to examine proteolytic digests in combination with NMR analysis purified sensitive, cryogenically cooled probe. Nano-liquid chromatography-tandem...

Summary The differences between archaeal and bacterial flagella are becoming more apparent as research on the structure progresses. One crucial difference is presence of a leader peptide preflagellins, which removed from flagellin prior to its incorporation into flagellar filament. enzyme responsible for removal was identified FlaK. FlaK Methanococcus voltae retains preflagellin peptidase activity when expressed in Escherichia coli used an vitro assay. Homologous recombination integration...

Archaeal flagellins are initially synthesized as preflagellins with a short, positively charged leader peptide, which is cleaved prior to the incorporation of mature into filament. While preflagellin peptidase activity had previously been detected in methanogen membranes, enzyme responsible for this not identified. We show here that FlaK Methanococcus maripaludis has activity. In an vitro assay, Escherichia coli membranes overexpressing voltae FlaB2 (as substrate) were combined E. M....

ABSTRACT Methanococcus voltae possesses four flagellin genes, two of which ( flaB1 and flaB2 ) have previously been reported to encode major components the flagellar filament. The remaining flaA flaB3 , are transcribed at lower levels, corresponding proteins remained undetected prior this work. Electron microscopy examination flagella isolated by detergent extraction whole cells revealed a curved, hook-like region varying length end long Enrichment curved resulted in identification FlaB3...

The correct delivery of noncytoplasmic proteins to locations both within and outside the cell depends on appropriate targeting signals. Protein translocation across bacterial plasma membrane eukaryal endoplasmic reticulum relies cleavable N-terminal signal peptides. Although peptides secreted in Bacteria Eukarya have been extensively studied at sequence, structure, functional levels, little is known nature archaeal In this report, genome-based analysis was performed an attempt define amino...

ABSTRACT The structure and function of two-component chemotactic signaling different aspects related to the motility bacteria archaea are key research areas in modern microbiology. Escherichia coli is traditional model organism used study chemotaxis motility. However, recent a wide range even some with lifestyles has provided new insight into ecophysiology chemotaxis, which essential for establishment pathogens or beneficial host. expanded organisms also permitted chemosensory pathways...

The outer membrane protein (OMP) TolC is the cell surface component of several drug efflux pumps that are responsible for bacterial resistance against a variety antibiotics. In this research, we investigated effects OMP on E. coli transport within saturated sands through column experiments using wild-type K12 strain (with TolC), as well corresponding transposon mutant (tolC::kan) and markerless deletion (ΔtolC). Our results showed could significantly enhance when ionic strength was 20 mM...

Pyomelanin is a brown/black extracellular pigment with antioxidant and iron acquisition properties that produced by number of different bacteria. Production pyomelanin in Pseudomonas aeruginosa contributes to increased resistance oxidative stress persistence chronic infections. We demonstrate production can be inhibited 2-[2-nitro-4-(trifluoromethyl) benzoyl]-1,3-cyclohexanedione (NTBC). This treatment increases sensitivity pyomelanogenic P. strains stress, without altering the growth rate...

ABSTRACT A soluble methyl-accepting chemotaxis protein (MCP) of Pseudomonas aeruginosa , McpS, showed polar localization by immunofluorescence microscopy. Overexpression McpS resulted in a dominant-negative effect on and caused loss clustering the general MCP population. The defines third, unexpected, paradigm for cellular localization.

The Pseudomonas aeruginosa Chp chemosensory system regulates twitching motility, intracellular adenosine 3′′ 5′-cyclic monophosphate (cAMP) levels and is postulated to be involved in directional towards phosphatidylethanolamine (PE). Because PilJ the only methyl-accepting chemotaxis protein (MCP) identified system, we determined role of mediating signal transduction for distinct outputs this system. Mutants that lack periplasmic domain (pilJΔ74-273) showed lower cAMP but retained PE. While...

Site-directed mutagenesis studies of the signal peptidase methanogenic archaeon Methanococcus voltae identified three conserved residues (Ser52, His122, and Asp148) critical for activity. The requirement one aspartic acid residue distinguishes archaeal enzyme from both Escherichia coli yeast Sec11 enzymes.

Pyomelanin is an extracellular red-brown pigment produced by several bacterial and fungal species. This derived from the tyrosine catabolism pathway contributes to increased oxidative stress resistance. production in Pseudomonas aeruginosa reduced a dose dependent manner through treatment with 2-[2-nitro-4-(trifluoromethyl)benzoyl]-1,3-cyclohexanedione (NTBC). We describe titration method using multiple concentrations of NTBC determine concentration drug that will reduce or abolish...

Pyomelanin is a reddish-brown pigment that provides bacteria and fungi protection from oxidative stress, reported to contribute infection persistence. Production of this can be inhibited by the anti-virulence agent 2-(2-nitro-4-trifluoromethylbenzoyl)-1,3-cyclohexanedione (NTBC). The Pseudomonas aeruginosa clinical isolate DKN343 exhibited high levels resistance NTBC, mechanism pyomelanin production in strain was uncharacterized. We determined due loss function homogentisate 1,2-dioxygenase...

In an attempt to improve upon a current mouse model of intestinal colonization by Escherichia coli O157:H7 used in this laboratory for vaccine development, nine clinical isolates the pathogen were screened their ability persist tract conventional adult CD-1 mice. None test E. capable colonizing these mice period more than two weeks. Most appeared be benign experimental host, but one isolate was lethal. This virulence correlated with latter produce large quantities Shiga-like toxin 2 vitro.

Archaeal flagellins are initially synthesized as preflagellins with a short, positively charged leader peptide, which is cleaved prior to the incorporation of mature into filament. While preflagellin peptidase activity had previously been detected in methanogen membranes, enzyme responsible for this not identified. We show here that FlaK Methanococcus maripaludis has activity. In an vitro assay, Escherichia coli membranes overexpressing voltae FlaB2 (as substrate) were combined E. M....

Pyomelanin is an extracellular red-brown pigment produced by several bacterial and fungal species. This derived from the tyrosine catabolism pathway contributes to increased oxidative stress resistance. production in Pseudomonas aeruginosa reduced a dose dependent manner through treatment with 2-[2-nitro-4-(trifluoromethyl)benzoyl]-1,3-cyclohexanedione (NTBC). We describe titration method using multiple concentrations of NTBC determine concentration drug that will reduce or abolish...

Abstract The recent characterization of partitioning proteins in the localization chemotaxis signal transduction systems was proposed to have broad implications for polarly-flagellated non-enterobacteriaceae gamma-proteobacteria. These studies showed that loss either protein resulted equivalent reductions swimming motility and inheritance. However, role these outside Vibrio spp . remains untested. Our on Pseudomonas aeruginosa revealed an unexpected ParP. While P. ParC ParP homologs are...