The aim and novelty of this paper are found in assessing the influence of inhibitors and antibiotics on intact cell MALDI-TOF mass spectra of the cyanobacterium Synechococcus sp. UPOC S4 and to check the impact on reliability of identification. Defining the limits of this method is important for its use in biology and applied science. The compounds included inhibitors of respiration, glycolysis, citrate cycle, and proteosynthesis. They were used at 1-10 μM concentrations and different periods of up to 3 weeks. Cells were also grown without inhibitors in a microgravity because of expected strong effects. Mass spectra were evaluated using controls and interpreted in terms of differential peaks and their assignment to protein sequences by mass. Antibiotics, azide, and bromopyruvate had the greatest impact. The spectral patterns were markedly altered after a prolonged incubation at higher concentrations, which precluded identification in the database of reference spectra. The incubation in microgravity showed a similar effect. These differences were evident in dendrograms constructed from the spectral data. Enzyme inhibitors affected the spectra to a smaller extent. This study shows that only a long-term presence of antibiotics and strong metabolic inhibitors in the medium at 10-5 M concentrations hinders the correct identification of cyanobacteria by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF).
- MeSH
- antibakteriální látky toxicita MeSH
- antimycin A analogy a deriváty toxicita MeSH
- azidy toxicita MeSH
- buněčné dýchání účinky léků MeSH
- chloramfenikol toxicita MeSH
- citrátový cyklus účinky léků MeSH
- deoxyglukosa toxicita MeSH
- fluoracetáty toxicita MeSH
- glykolýza účinky léků MeSH
- malonáty toxicita MeSH
- proteosyntéza účinky léků MeSH
- pyruváty toxicita MeSH
- reprodukovatelnost výsledků MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice metody MeSH
- stav beztíže MeSH
- streptomycin toxicita MeSH
- Synechococcus chemie účinky léků izolace a purifikace metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- enzymy * dějiny MeSH
- proteiny * dějiny MeSH
- Publikační typ
- biografie MeSH
- historické články MeSH
Cyanobacteria represent a bacterial phyllum characteristic by the ability to photosynthesize. They are potentially applicable for the production of useful compounds but may also cause poisoning or at least health problems as they can produce cyanotoxins. The introduction of a fast methodology is important not only for fundamental taxonomic purposes, but also for reliable identifications in biological studies. In this work, we have used matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of intact cells to study Chroococcidiopsis strains. A library of the obtained reference mass spectra containing characteristic peptide/protein profiles was examined by software tools to characterize similarities and differences applicable for diagnostics and taxonomy. Both a similarity tree and heat map constructed from the mass spectrometric data proved consistent with 16S rRNA sequencing results. We show as novelty that a binary matrix combining ferulic and sinapinic acids performs well in acquiring reproducible mass spectra of cyanobacteria. Using the matrix solvent, a protein extraction from cells was done. After polyacrylamide gel electrophoresis, the separated protein fractions were in-gel digested and the resulting peptides analyzed by liquid chromatography coupled with tandem mass spectrometry. For the first time, photosystem protein components, phycobilisome proteins, electron transport proteins, nitrogen-metabolism and nucleic acids binding-proteins, cytochromes plus other enzymes and various uncharacterized proteins could be assigned to characteristic peaks in the mass spectrometric profiles and some of them suggested as markers in addition to 30S and 50S ribosomal proteins known from previous studies employing intact cell mass spectrometry of microorganisms.
- MeSH
- bakteriální proteiny analýza izolace a purifikace MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- fylogeneze MeSH
- peptidy analýza izolace a purifikace MeSH
- RNA ribozomální 16S genetika MeSH
- sinice chemie klasifikace genetika MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice metody MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
This review reports on inhibitors of copper-containing amine oxidases and flavoprotein polyamine oxidases, which are structurally based on diamines. In the introduction, basic characteristics and classification of amine oxidases are described together with the significance of their synthetic inhibitors. The following text is divided into several chapters, which deal with diaminoketones, aza-diamines, unsaturated diamine analogs and diamines with heterocyclic substituents. Then it continues with diamine- and agmatine-based inhibitors of polyamine oxidases. Each chapter gives detailed information on the inhibition mode, potency and structural relationships. The conclusion points out possible roles of mechanism-based inhibitors of amine oxidases in physiological and medicinal research.
- MeSH
- agmatin farmakologie chemie MeSH
- aktivace enzymů fyziologie účinky léků MeSH
- diaminy farmakologie chemie MeSH
- financování organizované MeSH
- histaminasa antagonisté a inhibitory metabolismus MeSH
- inhibitory enzymů farmakologie chemie MeSH
- ketony farmakologie chemie MeSH
- lidé MeSH
- molekulární struktura MeSH
- oxidoreduktasy působící na CH-NH vazby antagonisté a inhibitory metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- přehledy MeSH
