Current standard methods for kinetic and genomic modeling cannot provide deep insight into metabolic regulation. Here, we developed and evaluated a multi-scale kinetic modeling approach applicable to any prokaryote. Specifically, we highlight the primary metabolism of the cyanobacterium Synechococcus elongatus PCC 7942. The model bridges metabolic data sets from cells grown at different CO2 conditions by integrating transcriptomic data and isozymes. Identification of the regulatory roles of isozymes allowed the calculation and explanation of the absolute metabolic concentration of 3-phosphoglycerate. To demonstrate that this method can characterize any isozyme, we determined the function of two glycolytic glyceraldehyde-3-phosphate dehydrogenases: one co-regulates high concentrations of the 3-phosphoglycerate, the other shifts the bifurcation point in hexose regulation, and both improve biomass production. Moreover, the regulatory roles of multiple phosphoglycolate phosphatases were defined for varying (non-steady) CO2 conditions, suggesting their protective role against toxic photorespiratory intermediates.
- MeSH
- adenosintrifosfát metabolismus MeSH
- bakteriální proteiny chemie fyziologie MeSH
- glyceraldehyd-3-fosfátdehydrogenasy chemie fyziologie MeSH
- homeostáza MeSH
- izoenzymy chemie fyziologie MeSH
- kinetika MeSH
- NADP metabolismus MeSH
- oxid uhličitý MeSH
- oxidace-redukce MeSH
- Synechococcus enzymologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The effect of protein kinase C (PKC) inhibitors on porcine oocyte activation by calcium ionophore A23187 was studied. Calcium ionophore applied in a 50 microM concentration for 10 min induced activation in 74% of oocytes matured in vitro. When the ionophore-treated oocytes were exposed to the effect of bisindolylmaleimide I, which inhibits calcium-dependent PKC isotypes (PKC-alpha, -beta(I), -beta(II), -gamma,) and calcium-independent PKC isotypes (PKC-delta, -epsilon), the portion of activated oocytes decreased (at a concentration of 100 nM, 2% of the oocytes were activated). Go6976, the inhibitor of calcium-dependent PKC isotypes PKC-alpha, -beta(I) did not prevent the action of the oocytes treated with calcium ionophore in concentrations from 1 to 100 microM. The inhibitor of PKC-beta(I) and beta(II) isotypes, hispidin, in a concentration of 2 microM-2 mM, was not effective either. The inhibitor of PKC-delta isotype, rottlerin, suppressed activation of the oocytes by calcium ionophore (no oocyte was activated at 10 microM concentration). The PKC-delta isotype in matured porcine oocytes, studied by Western blot analysis, appeared as non-truncated PKC-delta of 77.5 kDa molecular weight, on the one hand, and as truncated PKC-delta, which was present in the form of a doublet of approximately 62.5 and 68 kDa molecular weight, on the other hand. On the basis of these results, it can be supposed that PKC participates in the regulation of processes associated with oocyte activation. Calcium-dependent PKC-alpha, -beta isotypes do not seem to play any significant role in calcium activation. The activation seems to depend on the activity of the calcium-independent PKC-delta isoform.
- MeSH
- acetofenony farmakologie MeSH
- benzopyrany farmakologie MeSH
- calcimycin farmakologie MeSH
- indoly farmakologie MeSH
- inhibitory proteinkinas farmakologie MeSH
- ionofory farmakologie MeSH
- izoenzymy antagonisté a inhibitory fyziologie klasifikace MeSH
- karbazoly farmakologie MeSH
- maleimidy farmakologie MeSH
- oocyty fyziologie účinky léků MeSH
- prasata fyziologie MeSH
- proteinkinasa C antagonisté a inhibitory fyziologie klasifikace MeSH
- pyrony farmakologie MeSH
- vápník fyziologie MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- kazuistiky MeSH
- MeSH
- DNA gyráza analýza fyziologie MeSH
- izoenzymy analýza fyziologie MeSH
- karcinoid enzymologie MeSH
- lidé MeSH
- nádory plic enzymologie MeSH
- Check Tag
- lidé MeSH
- MeSH
- biologické modely MeSH
- cévní endotel fyziologie MeSH
- cévy fyziologie patologie zranění MeSH
- izoenzymy fyziologie izolace a purifikace MeSH
- myši MeSH
- plicní hypertenze etiologie patofyziologie MeSH
- plicní oběh fyziologie MeSH
- synthasa oxidu dusnatého izolace a purifikace klasifikace nedostatek MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- přehledy MeSH
- srovnávací studie MeSH
- MeSH
- ischemická choroba srdeční metabolismus patofyziologie MeSH
- ischemické přivykání škodlivé účinky MeSH
- izoenzymy enzymologie fyziologie MeSH
- krysa rodu rattus MeSH
- myokard enzymologie MeSH
- novorozená zvířata MeSH
- proteinkinasa C analýza fyziologie MeSH
- srdce patofyziologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
