Here, we report on the biochemical characterization of a new glycosylated bacteriocin (glycocin), ASM1, produced by Lactobacillus plantarum A-1 and analysis of the A-1 bacteriocinogenic genes. ASM1 is 43 amino acids in length with Ser18-O- and Cys43-S-linked N-acetylglucosamine moieties that are essential for its inhibitory activity. Its only close homologue, glycocin F (GccF), has five amino acid substitutions all residing in the flexible C-terminal 'tail' and a lower IC50 (0.9 nm) compared to that of ASM1 (1.5 nm). Asm/gcc genes share the same organization (asmH← →asmABCDE→F), and the asm genes reside on an 11 905-bp plasmid dedicated to ASM1 production. The A-1 genome also harbors a gene encoding a 'rare' bactofencin-type bacteriocin. As more examples of prokaryote S-GlcNAcylation are discovered, the functions of this modification may be understood.
- MeSH
- bakteriální geny genetika MeSH
- bakteriociny chemie genetika metabolismus MeSH
- fylogeneze MeSH
- glykosylace MeSH
- Lactobacillus plantarum chemie genetika MeSH
- novobiocin MeSH
- plazmidy genetika MeSH
- sekvence aminokyselin MeSH
- sekvence nukleotidů MeSH
- sekvenční analýza DNA MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
HIV protease (PR) is required for proteolytic maturation in the late phase of HIV replication and represents a prime therapeutic target. The regulation and kinetics of viral polyprotein processing and maturation are currently not understood in detail. Here we design, synthesize, validate and apply a potent, photodegradable HIV PR inhibitor to achieve synchronized induction of proteolysis. The compound exhibits subnanomolar inhibition in vitro. Its photolabile moiety is released on light irradiation, reducing the inhibitory potential by 4 orders of magnitude. We determine the structure of the PR-inhibitor complex, analyze its photolytic products, and show that the enzymatic activity of inhibited PR can be fully restored on inhibitor photolysis. We also demonstrate that proteolysis of immature HIV particles produced in the presence of the inhibitor can be rapidly triggered by light enabling thus to analyze the timing, regulation and spatial requirements of viral processing in real time.
- MeSH
- aminokumariny chemická syntéza farmakologie MeSH
- časové faktory MeSH
- fotolýza MeSH
- HEK293 buňky MeSH
- HIV-1 účinky léků fyziologie účinky záření MeSH
- HIV-proteasa chemie metabolismus MeSH
- inhibitory HIV-proteasy chemická syntéza farmakologie MeSH
- karbamáty chemická syntéza farmakologie MeSH
- kinetika MeSH
- lidé MeSH
- molekulární modely MeSH
- proteinové prekurzory antagonisté a inhibitory chemie metabolismus MeSH
- proteolýza účinky léků MeSH
- replikace viru MeSH
- světlo MeSH
- valin analogy a deriváty chemická syntéza farmakologie MeSH
- vazba proteinů MeSH
- vazebná místa MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Thirteen coagulase-negative, oxidase-negative, and novobiocin-susceptible staphylococci were isolated from human clinical specimens. The isolates were differentiated from known staphylococcal species on the basis of 16S rRNA, hsp60, rpoB, dnaJ, tuf, and gap gene sequencing, automated ribotyping, (GTG)5-PCR fingerprinting, and MALDI-TOF MS analysis. Phylogenetic analysis based on the 16S rRNA gene sequence indicated phylogenetic relatedness of the analyzed strains to Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus devriesei, and Staphylococcus lugdunensis. DNA-DNA hybridization experiments between representative strains CCM 8418(T), CCM 8421(T), and the closest phylogenetic neighbors confirmed that the isolates represent novel Staphylococcus species, for which the name Staphylococcus petrasii sp. nov. is proposed. Genotypic and phenotypic analyses unambiguously split the strains into two closely related subclusters. Based on the results, two novel subspecies S. petrasii subsp. petrasii subsp. nov. and S. petrasii subsp. croceilyticus subsp. nov. are proposed, with type strains CCM 8418(T) (=CCUG 62727(T)) and CCM 8421(T) (=CCUG 62728(T)), respectively.
- MeSH
- antibakteriální látky metabolismus MeSH
- bakteriální proteiny genetika MeSH
- DNA bakterií chemie genetika MeSH
- fylogeneze MeSH
- hybridizace nukleových kyselin MeSH
- koagulasa metabolismus MeSH
- lidé MeSH
- molekulární sekvence - údaje MeSH
- novobiocin metabolismus MeSH
- oxidoreduktasy metabolismus MeSH
- ribozomální DNA chemie genetika MeSH
- RNA ribozomální 16S genetika MeSH
- sekvenční analýza DNA MeSH
- shluková analýza MeSH
- stafylokokové infekce mikrobiologie MeSH
- Staphylococcus klasifikace genetika izolace a purifikace metabolismus MeSH
- techniky typizace bakterií MeSH
- ucho mikrobiologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Five isolates of coagulase-negative staphylococci were obtained from human urine, the gastrointestinal tract of squirrel monkeys, pig skin and from the environment. All key biochemical characteristics of the tested strains corresponded with the description of Staphylococcus xylosus species. However, partial 16S rRNA gene sequences obtained from analysed strains corresponded with those of Staphylococcus nepalensis reference strains, except for two strains which differed in one residue. Ribotyping with EcoRI and HindIII restriction enzymes, whole cell protein profile analysis performed by SDS-PAGE and SmaI macrorestriction analysis were used for more precise characterization and identification of the analysed strains. Obtained results showed that EcoRI and HindIII ribotyping and whole cell protein fingerprinting are suitable and reliable methods for the differentiation of S. nepalensis strains from the other novobiocin resistant staphylococci, whereas macrorestriction analysis was found to be a good tool for strain typing. The isolation of S. nepalensis is sporadic, and according to our best knowledge this study is the first report of the occurrence of this species in human clinical material as well as in other sources.
- MeSH
- DNA bakterií genetika chemie MeSH
- DNA fingerprinting MeSH
- financování organizované MeSH
- fylogeneze MeSH
- lidé MeSH
- novobiocin MeSH
- ribotypizace MeSH
- RNA ribozomální 16S analýza genetika MeSH
- sekvenční analýza DNA MeSH
- stafylokokové infekce mikrobiologie MeSH
- Staphylococcus fyziologie genetika izolace a purifikace klasifikace MeSH
- techniky typizace bakterií MeSH
- Check Tag
- lidé MeSH
A polyphasic identification approach was applied to a group of 11 novobiocin-resistant staphylococci isolated from human clinical materials. Phenotypic characteristics obtained by both commercial and conventional tests assigned eight strains as Staphylococcus xylosus and three strains as ambiguous S. xylosus/Staphylococcus equorum. In contrast to biotyping, ribotyping with EcoRI and HindIII restriction endonucleases and whole-cell protein fingerprinting assigned six analysed strains as S. equorum, and five strains as Staphylococcus succinus. Confirmation of the identification was done by partial 16S rRNA gene sequencing and S. equorum isolates were verified by a PCR assay targeting the sodA gene. From the data it has been implied that ribotyping and whole-cell protein analysis can be used to differentiate between the biochemically almost indistinguishable species S. xylosus, S. equorum and S. succinus. The present study confirms what is believed to be the first occurrence of S. equorum in a relevant human clinical material in the Czech Republic and describes what is believed to be the first-ever isolation of S. succinus from human clinical material.
- MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální léková rezistence MeSH
- bakteriální proteiny analýza genetika MeSH
- dítě MeSH
- DNA bakterií genetika chemie MeSH
- financování organizované MeSH
- kojenec MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- molekulární sekvence - údaje MeSH
- novobiocin farmakologie MeSH
- polymerázová řetězová reakce MeSH
- proteom analýza MeSH
- ribotypizace MeSH
- ribozomální DNA genetika chemie MeSH
- RNA ribozomální 16S genetika MeSH
- sekvenční analýza DNA MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- shluková analýza MeSH
- stafylokokové infekce mikrobiologie MeSH
- Staphylococcus fyziologie genetika izolace a purifikace klasifikace MeSH
- superoxiddismutasa genetika MeSH
- techniky typizace bakterií MeSH
- Check Tag
- dítě MeSH
- kojenec MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- MeSH
- antibiotická rezistence MeSH
- koagulasa MeSH
- kultivační média MeSH
- lidé MeSH
- novobiocin farmakologie MeSH
- stafylokokové infekce mikrobiologie MeSH
- Staphylococcus fyziologie klasifikace účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- přehledy MeSH
- MeSH
- fenotyp MeSH
- lidé MeSH
- mnohočetná léková rezistence MeSH
- novobiocin MeSH
- stafylokokové infekce mikrobiologie MeSH
- Staphylococcus klasifikace metabolismus účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- srovnávací studie MeSH
