A strain of Biatriospora sp. CCF 4378 was tested for the production of secondary metabolites under submerged fermentation conditions. Eleven compounds were isolated from the culture broth, and the structures of these compounds were determined using HRMS, NMR and X-ray analysis. In addition to six known naphthoquinone derivatives, i.e. ascomycone A, ascomycone B, 6-deoxyfusarubine, 6-deoxyanhydrofusarubine, herbarine and balticol A, one derivative of 2-azaanthraquinone, 6-deoxybostrycoidine, was also identified. Four new natural pyranonaphthoquinones were found, and these natural products were pleorubrin A, pleorubrin B, pleorubrin C and pleorubrin D. The toxicity on human cell lines of the crude naphthoquinone fraction and pure 6-deoxybostrycoidin, ascomycone B, pleorubrin B and 6-deoxyfusarubin was tested. Ascomycone B and 6-deoxyfusarubin elicited rapid cytotoxicity at micromolar concentrations.
- MeSH
- Ascomycota klasifikace genetika izolace a purifikace metabolismus MeSH
- buněčné linie MeSH
- endofyty klasifikace genetika izolace a purifikace metabolismus MeSH
- lidé MeSH
- molekulární struktura MeSH
- naftochinony chemie metabolismus farmakologie MeSH
- Ulmus mikrobiologie MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Four strains of the fungus Quambalaria cyanescens (Basidiomycota: Microstromatales), were used for the determination of secondary metabolites production and their antimicrobial and biological activities. A new naphthoquinone named quambalarine A, (S)-(+)-3-(5-ethyl-tetrahydrofuran-2-yliden)-5,7,8-trihydroxy-2-oxo-1,4-naphthoquinone (1), together with two known naphthoquinones, 3-hexanoyl-2,5,7,8-tetrahydroxy-1,4-naphthoquinone (named here as quambalarine B, 2) and mompain, 2,5,7,8-tetrahydroxy-1,4-naphthoquinone (3) were isolated. Their structures were determined by single-crystal X-ray diffraction crystallography, NMR and MS spectrometry. Quambalarine A (1) had a broad antifungal and antibacterial activity and is able inhibit growth of human pathogenic fungus Aspergillus fumigatus and fungi co-occurring with Q. cyanescens in bark beetle galleries including insect pathogenic species Beauveria bassiana. Quambalarine B (2) was active against several fungi and mompain mainly against bacteria. The biological activity against human-derived cell lines was selective towards mitochondria (2 and 3); after long-term incubation with 2, mitochondria were undetectable using a mitochondrial probe. A similar effect on mitochondria was observed also for environmental competitors of Q. cyanescens from the genus Geosmithia.
- MeSH
- antiinfekční látky chemie izolace a purifikace metabolismus farmakologie MeSH
- Basidiomycota metabolismus MeSH
- biologické přípravky chemie izolace a purifikace metabolismus farmakologie MeSH
- buněčné linie MeSH
- fermentace * MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A new polyene macrolide family, closely related to the pentaene macrolide antibiotic roflamycoin, was isolated from the both fermentation broth and biomass of Streptomyces durmitorensis wild-type strain MS405. The main compound was identified by NMR and Fourier transform ion cyclotron resonance mass spectrometry as 32,33-didehydroroflamycoin (1; DDHR). Additional four structurally related compounds were determined solely by MS analysis. DDHR induces cell death by apoptosis in various cancer cell lines as demonstrated by DNA fragmentation. Striking feature of DDHR is its internal fluorescence allowing visualization of labeled plasma membranes and internal membrane structures.
- MeSH
- barvení a značení MeSH
- buněčná membrána chemie metabolismus MeSH
- buněčná smrt účinky léků MeSH
- fluorescence MeSH
- hmotnostní spektrometrie MeSH
- lidé MeSH
- magnetická rezonanční spektroskopie MeSH
- makrolidy chemie metabolismus MeSH
- molekulární modely MeSH
- molekulární struktura MeSH
- nádorové buněčné linie MeSH
- polyeny chemie metabolismus MeSH
- protinádorové látky chemie metabolismus MeSH
- Streptomyces metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Lilac coloured species of Geosmithia lavendula produce a mixture of polyhydroxylated anthraquinones under condition of submerged fermentation. Three pigments had been isolated and identified earlier as a 1,3,6,8-tetrahydroxyanthraquinone (compound 7), rhodolamprometrin (1-acetyl 2,4,5,7-tetrahydroxyanthraquinone; compound 5), and 1-acetyl 2,4,5,7,8-penthahydroxyanthraquinone (compound 4). A new HPLC method was developed for the separation of three known and ten new anthraquinone pigments. In addition, five new pigments were determined by FTMS as coeluting impurities. The analyses were performed on a reversed phase column using gradient elution with a mobile phase system consisting of phosphate buffer (50 mM; pH=2.0) and acetonitrile. The structure evaluation was based namely on FTMS and UV-VIS spectrometry. The developed procedure was used for the determination of individual anthraquinones in fermentation broth of G. lavendula after 14 days of cultivation. The extractable amount and LOQ (both in μg ml(-1)) for the two main pigments from G. lavendula are 50.02 and 2.15 for compound 4, and 63.77 and 2.75, for compound 5, respectively.
- MeSH
- anthrachinony analýza chemie MeSH
- chromatografie s reverzní fází MeSH
- fermentace MeSH
- Hypocreales chemie MeSH
- lineární modely MeSH
- methanol MeSH
- reprodukovatelnost výsledků MeSH
- senzitivita a specificita MeSH
- tandemová hmotnostní spektrometrie metody MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Geosmithia fungi are little known symbionts of bark beetles. Secondary metabolites of lilac colored species G. lavendula and other nine Geosmithia species were investigated in order to elucidate their possible role in the interactions of the fungi with environment. Hydroxylated anthraquinones (yellow, orange, and red pigments), were found to be the most abundant compounds produced into the medium during the submerged cultivation. Three main compounds were identified as 1,3,6,8-tetrahydroxyanthraquinone (1), rhodolamprometrin (1-acetyl-2,4,5,7-tetrahydroxyanthraquinone; 2), and 1-acetyl-2,4,5,7,8-pentahydroxyanthraquinone (3). Compounds 2 and 3 (representing the majority of produced metabolites) inhibited the growth of G+-bacteria Staphylococcus aureus and Bacillus subtilis with minimum inhibitory concentration of 64-512 microg/mL. Anti-inflammatory activity detected as inhibition of cyclooxygenase-2 was found only for compound 3 at 1 and 10 microg/mL. Compound 2 interfered with the morphology, compound 3 with cell-cycle dynamics of adherent mammalian cell lines.
- MeSH
- anthrachinony farmakologie chemie metabolismus MeSH
- antibakteriální látky farmakologie chemie metabolismus MeSH
- antiflogistika nesteroidní farmakologie chemie metabolismus MeSH
- Bacillus subtilis účinky léků MeSH
- biologické pigmenty farmakologie chemie metabolismus MeSH
- biotechnologie metody MeSH
- buněčný cyklus účinky léků MeSH
- Ficus parazitologie MeSH
- financování organizované MeSH
- HeLa buňky MeSH
- hydroxylace MeSH
- Hypocreales metabolismus růst a vývoj MeSH
- inhibitory cyklooxygenasy farmakologie chemie metabolismus MeSH
- lidé MeSH
- nosatcovití fyziologie mikrobiologie MeSH
- spory hub metabolismus růst a vývoj MeSH
- Staphylococcus aureus účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
The structure of unique colorless needle crystals growing from the surface mycelium of the basidiomycete Quambalaria cyanescens and identified as (+)-globulol was followed by mass spectrometry, X-ray diffraction, and polarimetry. The mechanism of (+)-globulol fragmentation is proposed based on collision induced dissociation mass spectrometry. X-Ray analysis revealed that crystal packing is governed by hydrogen bond O-H.....O connecting the molecules into an infinite helix along a 3-fold screw axis propagating along the longest dimension of the needle crystal (c-axis of the unit cell). The X-ray diffraction data correspond well with the proposed structure determined by mass spectrometry.
- MeSH
- Basidiomycota chemie izolace a purifikace metabolismus MeSH
- brouci mikrobiologie MeSH
- difrakce rentgenového záření MeSH
- hmotnostní spektrometrie MeSH
- krystalizace MeSH
- molekulární struktura MeSH
- mycelium chemie metabolismus MeSH
- seskviterpeny chemie metabolismus MeSH
- vodíková vazba MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
Peptides eluted from peripheral blood cells of HLA-B*2705 healthy donor were analyzed by LC MALDI MS/MS and LC ESI FTMS techniques. The sequences of 92 peptide ligands identified from one healthy blood donor by LC MALDI-TOF MS/MS were compared with those previously published from in vitro long-term cell cultures available in SYFPEITHI database and splenocytes. It was found that 18 sequences confirmed within 1ppm mass error by LC ESI FTMS were already described and 3 of them matched with those previously reported from HLA-B*2705 splenocytes. Another 38 sequences validated within the same mass error were not found in SYFPEITHI database and are identified here for the first time. Finally, 36 sequences (5 sequences already published in SYFPEITHI database) were evaluated by LC MALDI-TOF MS/MS but no matches in the list of monoisotopic masses obtained from LC ESI FTMS were found.
- MeSH
- alkoholoxidoreduktasy MeSH
- ankylózující spondylitida genetika metabolismus MeSH
- autoimunita genetika MeSH
- databáze proteinů MeSH
- dospělí MeSH
- endopeptidasy metabolismus MeSH
- financování organizované MeSH
- genetická predispozice k nemoci MeSH
- histony genetika metabolismus MeSH
- HLA-B27 antigen analýza imunologie izolace a purifikace MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
- interakční proteinové domény a motivy MeSH
- krevní buňky imunologie metabolismus MeSH
- lidé MeSH
- peptidové mapování MeSH
- peptidy analýza imunologie izolace a purifikace MeSH
- proteiny tepelného šoku HSC70 genetika imunologie krev MeSH
- proteiny vázající GTP - alfa-podjednotky Gs genetika imunologie krev MeSH
- sekvenční seřazení MeSH
- software MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- srovnávací studie MeSH
HLA-B27 is a relative risk factor for ankylosing spondylitis (AS) and is present in about 10% in European populations but in 95% of AS patients. Various data suggest that the HLA-B27 molecule itself could be the strongest risk factor, but there is no explanation for this association. To define differential antigen presenting features of HLA-B27 in healthy individuals and AS patients, a question that cannot be addressed by biochemical studies on cell lines, the HLA-B27 protein was purified from peripheral blood lymphocytes of AS patients and healthy controls and pool sequencing of the bound peptides was performed. Results show that peptides are rich in proline (Pro) and the content of arginine (Arg) is much lower in comparison with sequences listed in the register of peptides eluted from cell cultures. Statistically significant differences were detected in frequencies of a subset of amino acids, predominantly at positions in the middle of the peptides. The frequency of Glu was increased and Gln was decreased in peptides from AS patients. Detailed analysis of purity of the immunoisolated HLA molecules excluded that the peptides might originate from any co-purified HLA molecules other than B27. We conclude that statistically significant increase in the Glu/Gln ratio of peptides from AS patients, consistent with increased deamidation in vivo, may account for differential antigenicity of HLA-B27 in patients. Source protein(s) of deamidated peptides remain unknown.
- MeSH
- ankylózující spondylitida genetika MeSH
- arginin genetika MeSH
- dospělí MeSH
- financování organizované MeSH
- glutamin genetika MeSH
- HLA-B27 antigen genetika chemie MeSH
- kyselina glutamová genetika MeSH
- lidé MeSH
- peptidové fragmenty genetika chemie genetika MeSH
- prolin genetika MeSH
- rizikové faktory MeSH
- sekvenční analýza proteinů MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- srovnávací studie MeSH
The sequences and profiles of peptides which bind to HLA-B*2705 splenocytes and peripheral blood cells were compared with those previously published from in vitro long-term cell cultures. B*2705 peptide profile analysed by solid-phase Edman degradation and 15 individual peptide sequences determined by LC-MS/MS were partially similar to those defined from in vitro long-term cell cultures. Arg at P2 was found in 11 of 15 sequenced peptides (73.3%). This value is lower in comparison with other published data. Two sequences were matching to unknown proteins, which displayed similarity with myosin. These are first data on peptide sequences isolated directly from HLA-B27 molecules without prior in vitro propagation of the cells.
