Hydrophilic interaction liquid chromatography (HILIC) connected with electrospray high-resolution tandem mass spectrometry (MS) was used for the analysis of unusual amino acid (AA) substituted phosphatidylglycerols (PG) and cardiolipins (CL) in mesophilic and thermophilic bacteria. Individual peaks from the lipid class separation by HILIC were isolated and hydrolyzed to determine the absolute configuration of the aminoacyl side chain. The configuration of the aminoacyl side chain was assigned by indirect liquid chromatography (LC) enantiomer separation after the hydrolysis of the aminoacylated (aminoacyl) lipids using N-(4-nitrophenoxycarbonyl)-l-phenylalanine 2-methoxyethyl ester as chiral derivatizing agent and reversed phase LC-MS for analysis. When two chromatographic methods were combined, less common AAs, such as d-allo-Ile and d-allo-Thr, were identified. The taxonomic classification of bacteria showed that bacteria of the family Bacillaceae (Bacillus and Geobacillus) produce branched-chain AAs, that is, d-allo-Ile, d-Ile, and d-Leu. These AAs were present only in the genera Bacillus and Geobacillus and not in Alicyclobacillus acidoterrestris (family Alicyclobacillaceae). On the contrary, hydroxy AAs, that is, l- and d-Thr, and l- and d-allo-Thr, were identified as aminoacyl-PG and aminoacyl-CL in A. acidoterrestris and were not present in the genera Bacillus and Geobacillus. Therefore, the complete analysis made it possible to identify the stereochemistry of AAs in aminoacyl PGs and CLs and use this fact for chemotaxonomy.

• Klíčová slova
• HILIC, achiral chromatography, amino acids, aminoacylated cardiolipins, aminoacylated phosphatidylglycerols,
• MeSH
• aminokyseliny chemie   analýza   MeSH
• Bacteria chemie   MeSH
• chromatografie kapalinová metody   MeSH
• fosfatidylglyceroly * chemie   MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací metody   MeSH
• hydrofobní a hydrofilní interakce MeSH
• kardiolipiny * chemie   analýza   MeSH
• stereoizomerie MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aminokyseliny MeSH
• fosfatidylglyceroly * MeSH
• kardiolipiny * MeSH

Unusual glucose-substituted cardiolipins (Glcx-CLs) in three genera of thermophilic bacteria, having more than one glycosidically linked glucose to the hydroxyl of the central glycerol of Glcx-CLs were identified for the first time in thermophilic bacteria of the genera Geobacillus, Meiothermus, and Thermus. The number of glucoses reached up to five units. The structure of glycosidically linked oligosaccharides was determined based on shotgun analysis MS (electrospray high-resolution tandem mass spectrometry), partially methylated alditol acetates were identified by GC-MS, both electron ionization (EI) and positive chemical ionization (PCI), hydrophilic interaction liquid chromatography (HILIC) separation and identification of CLs glycosides by high resolution MS-ESI, and digestion by specific glycosidases.

• Klíčová slova
• Electrospray mass spectrometry, Glucosyl-cardiolipins, Hydrophilic interaction liquid chromatography, Partially methylated alditol acetates, Thermophilic bacteria,
• MeSH
• Bacteria MeSH
• chromatografie kapalinová metody   MeSH
• glukosa MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací metody   MeSH
• kardiolipiny * analýza   MeSH
• plynová chromatografie s hmotnostně spektrometrickou detekcí MeSH
• tandemová hmotnostní spektrometrie * metody   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• glukosa MeSH
• kardiolipiny * MeSH

High resolution electrospray ionization tandem mass spectrometry (HR-ESI-MS/MS) was used to analyze cardiolipins (Ptd2Gro) including their plasmalogen forms from three species of the anaerobic beer-spoilage bacterial genus Pectinatus. Cardiolipins including their plasmalogens were analyzed by HR-ESI-MS/MS on Orbitrap and almost 100 cardiolipins (i.e. tetra-acyl--Ptd2Gro, plasmenyl-tri-acyl--PlsPtd2Gro, and di-plasmenyl-di-acyl--Pls2Ptd2Gro) of three classes were identified. The structures of the molecular species that consist of various regioisomers and structurally similar compounds were revealed in detail. The high resolution mass spectrometry allowed the unambiguous structural assignment of Ptd2Gro, PlsPtd2Gro, and Pls2Ptd2Gro in the three species of Pectinatus, which contain predominantly odd numbered fatty acids.

• MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací metody   MeSH
• kardiolipiny analýza   chemie   MeSH
• kontaminace potravin MeSH
• molekulární struktura MeSH
• Pectinatus chemie   MeSH
• pivo mikrobiologie   MeSH
• plasmalogeny analýza   chemie   MeSH
• potravinářská mikrobiologie MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• kardiolipiny MeSH
• plasmalogeny MeSH

The rational design of molecules with selective intracellular targeting is a great challenge for contemporary chemistry and life sciences. Here, we demonstrate a rational approach to development of compartment-specific fluorescent dyes from the γ-aryl substituted pentamethine family. These novel dyes exhibit an extraordinary affinity and selectivity for cardiolipin in inner mitochondrial membrane and possess excellent photostability, fluorescent properties, and low phototoxicity. Selective imaging of live and fixed mitochondria was achieved in various cell lines using nanomolar concentrations of these dyes. Their high localization specificity and low toxicity enables study of morphological changes, structural complexity, and dynamics of mitochondria playing a pivotal role in many pathological diseases. These far-red emitting dyes could also serve in a variety of biomedical applications.

• MeSH
• buněčné linie MeSH
• DNA analýza   MeSH
• fluorescenční barviva analýza   metabolismus   MeSH
• kardiolipiny analýza   metabolismus   MeSH
• krystalografie rentgenová MeSH
• kultivované buňky MeSH
• kur domácí MeSH
• lidé MeSH
• ligandy MeSH
• mitochondrie metabolismus   ultrastruktura   MeSH
• molekulární modely MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA MeSH
• fluorescenční barviva MeSH
• kardiolipiny MeSH
• ligandy MeSH

We used direct ESI-MS analysis to identify derivatives of cardiolipin molecular species (i.e. O-acyl glycosylated cardiolipins) from the thermophilic bacterium Alicyclobacillus acidoterrestris. We used triple-quadrupole type mass spectrometer for analysis of this complex lipid and enzymatic hydrolysis and 1H and 13C NMR for the identification of these cardiolipin derivatives. These techniques enabled us to identify and quantify the specific molecular species profiles of derivatives of cardiolipin directly from lipid extracts of the bacterium including the identification of the sugar moiety as alpha-D-mannose and all five acyls including their positional isomers.

• MeSH
• Bacteria chemie   MeSH
• glykosylace MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací metody   MeSH
• hmotnostní spektrometrie metody   MeSH
• izotopy kyslíku chemie   MeSH
• kardiolipiny analýza   izolace a purifikace   MeSH
• klinické laboratorní techniky MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• izotopy kyslíku MeSH
• kardiolipiny MeSH