Colabomycin E is a new member of the manumycin-type metabolites produced by the strain Streptomyces aureus SOK1/5-04 and identified by genetic screening from a library of streptomycete strains. The structures of colabomycin E and accompanying congeners were resolved. The entire biosynthetic gene cluster was cloned and expressed in Streptomyces lividans. Bioinformatic analysis and mutagenic studies identified components of the biosynthetic pathway that are involved in the formation of both polyketide chains. Recombinant polyketide synthases (PKSs) assembled from the components of colabomycin E and asukamycin biosynthetic routes catalyzing the biosynthesis of "lower" carbon chains were constructed and expressed in S. aureus SOK1/5-04 ΔcolC11-14 deletion mutant. Analysis of the metabolites produced by recombinant strains provided evidence that in both biosynthetic pathways the length of the lower carbon chain is controlled by an unusual chain-length factor supporting biosynthesis either of a triketide in asukamycin or of a tetraketide in colabomycin E. Biological activity assays indicated that colabomycin E significantly inhibited IL-1β release from THP-1 cells and might thus potentially act as an anti-inflammatory agent.
- MeSH
- antiflogistika chemie metabolismus farmakologie MeSH
- bicyklické sloučeniny heterocyklické chemie metabolismus farmakologie MeSH
- buněčné linie MeSH
- interleukin-1beta sekrece MeSH
- lidé MeSH
- molekulární struktura MeSH
- polynenasycené alkamidy chemie metabolismus farmakologie MeSH
- Streptomyces chemie metabolismus MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- Escherichia coli imunologie izolace a purifikace MeSH
- interleukin-1beta sekrece MeSH
- Lactobacillus casei imunologie izolace a purifikace MeSH
- oxid dusnatý sekrece MeSH
- probiotika * farmakologie metabolismus terapeutické užití MeSH
- TNF-alfa sekrece MeSH
- ultrafiltrace metody přístrojové vybavení MeSH
- Publikační typ
- práce podpořená grantem MeSH
Kinetin (N6-furfuryladenine) belongs to a group of plant growth hormones involved in cell division, differentiation and other physiological processes. One of the possible ways to obtain biologically active compounds is to complex biologically relevant natural compounds to suitable metal atoms. In this work, two structural groups of Zn(II) complexes [Zn(L(n))2Cl2]·Solv (1-5) and [Zn(HL(n))Cl3] · xL(n) (6-7); n=1-5, Solv=CH3OH for 1 and 2H2O for 2; x =1 for 6 and 2 for 7; involving a phytohormone kinetin and its derivatives (L(n)) were evaluated for their ability to modulate secretion of tumour necrosis factor (TNF)-α, interleukin (IL)-1β and matrix metalloproteinase (MMP)-2 in a lipopolysaccharide (LPS)-activated macrophage-like THP-1 cell model. The penetration of the complexes to cells was also detected. The mechanism of interactions of the zinc(II) complexes with a fluorescent sensor N-(6-methoxy-8-quinolyl)-p-toluene sulphonamide (TSQ) and sulfur-containing biomolecules (l-cysteine and reduced glutathione) was studied by electrospray-ionization mass spectrometry and flow-injection analysis with fluorescence detection. The present study showed that the tested complexes exhibited a low cytotoxic effect on the THP-1 cell line (IC50>40 µM), apart from complex 4, with an IC50=10.9 ± 1.1 µM. Regarding the inflammation-related processes, the Zn(II) complexes significantly decreased IL-1β production by a factor of 1.47-2.22 compared with the control (DMSO), but did not affect TNF-α and MMP-2 secretions. However, application of the Zn(II) complexes noticeably changed the pro-MMP-2/MMP-2 ratio towards a higher amount of maturated MMP-2, when they induced a 4-times higher production of maturated MMP-2 in comparison with the vehicle-treated cells under LPS stimulation. These results indicated that the complexes are able to modulate an inflammatory response by influencing secretion and activity of several inflammation-related cytokines and enzymes.
- MeSH
- aktivace makrofágů účinky léků MeSH
- aminochinoliny MeSH
- antiflogistika chemická syntéza farmakologie MeSH
- biologický transport MeSH
- chloridy chemie MeSH
- cystein chemie MeSH
- exprese genu účinky léků MeSH
- fluorescenční barviva MeSH
- glutathion chemie MeSH
- interleukin-1beta antagonisté a inhibitory genetika sekrece MeSH
- kationty dvojmocné MeSH
- kinetin chemie MeSH
- komplexní sloučeniny chemická syntéza farmakologie MeSH
- lidé MeSH
- lipopolysacharidy farmakologie MeSH
- makrofágy cytologie účinky léků sekrece MeSH
- matrixová metaloproteinasa 2 genetika sekrece MeSH
- nádorové buněčné linie MeSH
- TNF-alfa genetika sekrece MeSH
- tosylové sloučeniny MeSH
- viabilita buněk účinky léků MeSH
- zinek chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
