Lipophosphonoxins (LPPOs) are small modular synthetic antibacterial compounds that target the cytoplasmic membrane. First-generation LPPOs (LPPO I) exhibit an antimicrobial activity against Gram-positive bacteria; however they do not exhibit any activity against Gram-negatives. Second-generation LPPOs (LPPO II) also exhibit broadened activity against Gram-negatives. We investigated the reasons behind this different susceptibility of bacteria to the two generations of LPPOs using model membranes and the living model bacteria Bacillus subtilis and Escherichia coli. We show that both generations of LPPOs form oligomeric conductive pores and permeabilize the bacterial membrane of sensitive cells. LPPO activity is not affected by the value of the target membrane potential, and thus they are also active against persister cells. The insensitivity of Gram-negative bacteria to LPPO I is probably caused by the barrier function of the outer membrane with LPS. LPPO I is almost incapable of overcoming the outer membrane in living cells, and the presence of LPS in liposomes substantially reduces their activity. Further, the antimicrobial activity of LPPO is also influenced by the phospholipid composition of the target membrane. A higher proportion of phospholipids with neutral charge such as phosphatidylethanolamine or phosphatidylcholine reduces the LPPO permeabilizing potential.
- MeSH
- antibakteriální látky chemická syntéza farmakologie MeSH
- Bacillus subtilis chemie cytologie účinky léků MeSH
- Escherichia coli chemie cytologie účinky léků MeSH
- fosfatidylcholiny analýza metabolismus MeSH
- fosfatidylethanolaminy analýza metabolismus MeSH
- kationické antimikrobiální peptidy chemická syntéza farmakologie MeSH
- lipidové dvojvrstvy MeSH
- membránové potenciály účinky léků MeSH
- mikrobiální testy citlivosti MeSH
- permeabilita buněčné membrány MeSH
- vnější bakteriální membrána chemie účinky léků metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The comprehensive approach for the lipidomic characterization of human breast cancer and surrounding normal tissues is based on hydrophilic interaction liquid chromatography (HILIC)-electrospray ionization mass spectrometry (ESI-MS) quantitation of polar lipid classes of total lipid extracts followed by multivariate data analysis using unsupervised principal component analysis (PCA) and supervised orthogonal partial least square (OPLS). This analytical methodology is applied for the detailed lipidomic characterization of ten patients with the goal to find the statistically relevant differences between tumor and normal tissues. This strategy is selected for better visualization of differences, because the breast cancer tissue is compared with the surrounding healthy tissue of the same patient, therefore changes in the lipidome are caused predominantly by the tumor growth. A large increase of total concentrations for several lipid classes is observed, including phosphatidylinositols, phosphatidylethanolamines, phosphatidylcholines, and lysophosphatidylcholines. Concentrations of individual lipid species inside the abovementioned classes are also changed, and in some cases, these differences are statistically significant. PCA and OPLS analyses enable a clear differentiation of tumor and normal tissues based on changes of their lipidome. A notable decrease of relative abundances of ether and vinylether (plasmalogen) lipid species is detected for phosphatidylethanolamines, but no difference is apparent for phosphatidylcholines.
- MeSH
- analýza hlavních komponent MeSH
- chromatografie kapalinová metody MeSH
- fosfatidylcholiny analýza MeSH
- fosfatidylethanolaminy analýza MeSH
- fosfatidylinositoly analýza MeSH
- fosfolipidy analýza chemie MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací metody MeSH
- hydrofobní a hydrofilní interakce MeSH
- lidé MeSH
- lipidy analýza chemie MeSH
- lysofosfatidylcholiny analýza MeSH
- multivariační analýza MeSH
- nádory prsu metabolismus patologie MeSH
- referenční hodnoty MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Lipids form a significant part of animal organs and they are responsible for important biological functions, such as semi-permeability and fluidity of membranes, signaling activity, anti-inflammatory processes, etc. We have performed a comprehensive nontargeted lipidomic characterization of porcine brain, heart, kidney, liver, lung, spinal cord, spleen, and stomach using hydrophilic interaction liquid chromatography (HILIC) coupled to electrospray ionization mass spectrometry (ESI/MS) to describe the representation of individual lipid classes in these organs. Detailed information on identified lipid species inside classes are obtained based on relative abundances of deprotonated molecules [M-H](-) in the negative-ion ESI mass spectra, which provides important knowledge on phosphatidylethanolamines and their different forms of fatty acyl linkage (ethers and plasmalogens), phosphatidylinositols, and hexosylceramides containing nonhydroxy- and hydroxy-fatty acyls. The detailed analysis of identified lipid classes using reversed-phase liquid chromatography in the second dimension was performed for porcine brain to determine more than 160 individual lipid species containing attached fatty acyls of different acyl chain length, double-bond number, and positions on the glycerol skeleton. The fatty acid composition of porcine organs is determined by gas chromatography with flame ionization detection after the transesterification with sodium methoxide.
- MeSH
- chromatografie kapalinová metody MeSH
- chromatografie plynová metody MeSH
- fosfatidylethanolaminy analýza MeSH
- fosfolipidy analýza MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací metody MeSH
- hydrofobní a hydrofilní interakce MeSH
- játra chemie MeSH
- ledviny chemie MeSH
- lipidy analýza chemie MeSH
- mastné kyseliny analýza MeSH
- mícha chemie MeSH
- mozek - chemie MeSH
- myokard chemie MeSH
- plasmalogeny analýza MeSH
- plíce chemie MeSH
- prasata MeSH
- slezina chemie MeSH
- žaludek chemie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Phospholipid (PL) and fatty acid (FA) compositions of the plasma membrane (PM), as well as the FA composition of the PM phosphatidylcholine (PtdCho) and phosphatidylethanolamine (PtdEtn) in the pure culture (zero generation) and the first three recycled generations of the bottom-fermenting brewer's yeast, have been determined. The PL composition differed markedly among the generations; in the zero generation, phosphatidylinositol (PtdIns) was the main PL, accounting for 27% of total PLs, followed by phosphatidic acid and PtdCho. In all recycled generations, the main PL was PtdCho with a marked increase in the first generation compared with the zero (32% and 20%, respectively), followed by PtdIns in the first and second generations. In the FA composition of the PM, 22 FAs were identified, ranging from C(10) to C(26). The compositions of the PM FAs, as well as those of PtdCho and PtdEtn, were characterised by a high preponderance of C(16) acids. Saturated FAs prevailed in the zero generation, whilst unsaturated prevailed in the first and second generation. Although the profiles of FAs in PtdCho and PtdEtn were similar, some marked differences were observed, pointing out to their specific functions in the regulation of membrane properties.
- MeSH
- buněčná membrána chemie fyziologie MeSH
- fermentace MeSH
- fosfatidylcholiny analýza MeSH
- fosfatidylethanolaminy analýza MeSH
- fosfolipidy analýza MeSH
- fyziologický stres fyziologie MeSH
- mastné kyseliny analýza MeSH
- Saccharomyces cerevisiae chemie fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
