Structure and absolute configuration of an unsaturated anteiso fatty acid from Bacillus megaterium.

Gas chromatography in combination with electron ionization mass spectrometry (GC/EI-MS) was used to determine the fatty acids of a membrane lipid from Bacillus megaterium. Special attention was put on the structure and absolute configuration of a monoenoic fatty acid previously described in this sample. GC/EI-MS operated in the selected ion monitoring mode was used to determine twelve fatty acids in the bacterium.

Effects of lipid composition on the membrane activity and lipid phase behaviour of Vibrio sp. DSM14379 cells grown at various NaCl concentrations.

The membrane lipid composition of living cells generally adjusts to the prevailing environmental and physiological conditions. In this study, membrane activity and lipid composition of the Gram-negative bacterium Vibrio sp. DSM14379, grown aerobically in a peptone-yeast extract medium supplemented with 0.

Lipid dependence and activity control of phosphatidylserine synthase from Escherichia coli.

The activity of phosphatidylserine synthase from Escherichia coli depends significantly on the nature and level of the lipids in the matrix, at which the enzyme is operating. To elucidate the role of anionic lipids in the regulation of PtdSer synthase, its activity was studied in mixed micelles containing phosphatidylglycerol (one charge) or diphosphatidylglycerol (two charges), the two main anionic membrane lipids in E. coli.

Regulation of lipid composition in Acholeplasma laidlawii and Escherichia coli membranes: NMR studies of lipid lateral diffusion at different growth temperatures.

Lipid lateral diffusion coefficients have been directly determined by pulsed field gradient NMR spectroscopy on macroscopically aligned, fully hydrated lamellar phases containing dimyristoylphosphatidylcholine and total lipid extracts from Acholeplasma laidlawii and Escherichia coli. The temperature dependence of the diffusion coefficient was of the Arrhenius type in the temperature interval studied. The sharp increase in the diffusion coefficient at the growth temperature of E.

The physico-chemical characteristics of the phosphocholine-containing glycoglycerolipid MfGL-II govern the permeability properties of Mycoplasma fermentans.

Mycoplasma fermentans seems to be involved in several pathogenic conditions in humans, and is among other things capable of fusing with T-cells and lymphocytes. The choline-containing phosphoglycolipid 6'-O-(3"-phosphocholine-2"-amino-1"-phospho-1",3"-propanediol)-alpha-D-glucopyranosyl-(1'-->3)-1,2-diacylglycerol (MfGL-II) in the membrane of M. fermentans has been suggested to enhance the fusion process, and the characteristics of MfGL-II were therefore investigated.

alpha-methylene ordering of acyl chains differs in glucolipids and phosphatidylglycerol from Acholeplasma laidlawii membranes: (2)H-NMR quadrupole splittings from individual lipids in mixed bilayers.

A Acholeplasma laidlawii strain A-EF22 was grown in a medium supplemented with alpha-deuterated oleic acid. Phosphatidylglycerol (PG), the glucolipids monoglucosyldiacylglycerol (MGlcDAG), diglucosyldiacylglycerol (DGlcDAG) and monoacyldiglucosyldiacylglycerol, and the phosphoglucolipid glycerophosphoryldiglucosyldiacylglycerol (GPDGlcDAG) were purified, and the phase behaviour and molecular ordering for the individual lipids, as well as for mixtures of the lipids, were studied by (2)H-, (31)P-NMR and X-ray scattering methods. The chemical structure of all the A.

Interaction of phosphatidylserine synthase from E. coli with lipid bilayers: coupled plasmon-waveguide resonance spectroscopy studies.

The interaction of phosphatidylserine (PS) synthase from Escherichia coli with lipid membranes was studied with a recently developed variant of the surface plasmon resonance technique, referred to as coupled plasmon-waveguide resonance spectroscopy. The features of the new technique are increased sensitivity and spectral resolution, and a unique ability to directly measure the structural anisotropy of lipid and proteolipid films. Solid-supported lipid bilayers with the following compositions were used: 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC); POPC-1-palmitoyl-2-oleoyl-sn-glycero-3-phosphate (POPA) (80:20, mol/mol); POPC-POPA (60:40, mol/mol); and POPC-1-palmitoyl-2-oleoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (POPG) (75:25, mol/mol).

Reconstituted phosphatidylserine synthase from Escherichia coli is activated by anionic phospholipids and micelle-forming amphiphiles.

The activity of phosphatidylserine (PS) synthase (CDP-1, 2-diacyl-sn-glycerol: l-serine O-phosphatidyltransferase, EC 2.7.8.

Total lipids with short and long acyl chains from Acholeplasma form nonlamellar phases.

The cell-wall-less bacterium Acholeplasma laidlawii A-EF22 synthesizes eight glycerolipids. Some of them form lamellar phases, whereas others are able to form normal or reversed nonlamellar phases. In this study we examined the phase properties of total lipid extracts with limiting average acyl chain lengths of 15 and 19 carbon atoms.

Lipids in total extracts from Acholeplasma laidlawii A pack more closely than the individual lipids. Monolayers studied at the air-water interface.

Pressure-area curves were obtained at 25, 35 and 45 degrees C for total lipid extracts and four individual glucolipids isolated from Acholeplasma laidlawii strain A-EF22. The glucolipids are 1,2-diacyl-3-0-(alpha-D-glucopyranosyl)-sn-glycerol (MGlcDAG), 1,2 -diacyl-3-0-[alpha-D-glucopyranosyl-(1-->2)-0-alpha-D-glucopyranosyl] -sn-glycerol (DGlcDAG), 1,2-diacyl-3-0-[alpha-D-glucopyranosyl-(1-->2)-0-(6-0-acyl-alpha-D-gluco pyranosyl)]-sn-glycerol (MADGlcDAG), and 1,2-diacyl-3-0-[glycerophosphoryl-6-0-(alpha-D-glucopyranosyl-(1-- >)-0-alpha-D-glucopyranosyl)]-sn-glycerol (GPDGlcDAG). The total lipid extracts were obtained from A.

Occurrence of monoacyl-diglucosyl-diacyl-glycerol and monoacyl-bis-glycerophosphoryl-diglucosyl-diacyl-glycerol in membranes of Acholeplasma laidlawii strain B-PG9.

It is shown by thin-layer and high-performance liquid chromatography that the two membrane lipids monoacyl-diglucosyl-diacyl-glycerol (MADGlcDAG) and monoacyl-bis-glycerophosphoryl-diglucosyl-diacyl-glycerol are synthesized by Acholeplasma laidlawii strain B-PG9 when the cells are grown in two different growth media. The two lipids are also synthesized by A. laidlawii strain A-EF22 and their chemical structures have been determined previously by NMR spectroscopy.

Cryo-TEM and NMR studies of a micelle-forming phosphoglucolipid from membranes of Acholeplasma laidlawii A and B.

The chemical structure of a phosphoglucolipid from the membrane of the bacterium Acholeplasma laidlawii strain B-PG9 has been determined by high resolution NMR to be 1,2-diacyl-3-O-[glycerophosphoryl-6-O-(alpha-D-glucopyranosyl-(1 -->2)-O-alpha-D-glucopyranosyl)]-sn-glycerol (GPDGlcDAG). It was concluded that this lipid has exactly the same structure as one of the phosphoglucolipids from A. laidlawii strain A-EF22.

Two-dimensional 1H-NMR of transmembrane peptides from Escherichia coli phosphatidylglycerophosphate synthase in micelles.

Two 28-residue peptides, PTLLTLFRVILIPFFVLVFYKKKGKKKG [Pgs-(6-25)-peptidyl-KKKGKKKG; Pgs peptide A] and VEYAGIALFFVAAVLTLWSMLQYLSAAR [Pgs-(149-176)-peptide, Pgs peptide E], were synthesized and studied by CD and two-dimensional 1H-NMR spectroscopy. The first 20 amino acid residues of Pgs peptide A are identical to one predicted transmembrane segment (Pro6-Tyr25) of the integral membrane protein phosphatidylglycerophosphate synthase (Pgs) of Escherichia coli. Pgs peptide E is identical to another predicted transmembrane segment (Val149-Arg176), which is located in the C-terminal end of this lipid synthase.

New aspects on membrane lipid regulation in Acholeplasma laidlawii A and phase equilibria of monoacyldiglucosyldiacylglycerol.

A new membrane lipid, monoacyldiglucosyldiacylglycerol (MADGlcDAG), was recently discovered in Acholeplasma laidlawii strain A-EF22, demanding a new study of the biosynthetic regulation, and the phase behavior, of the glucolipids in this organism. The only liquid-crystalline phase formed by MADGlcDAG is a reversed hexagonal phase. A.

Wild-type Escherichia coli cells regulate the membrane lipid composition in a "window" between gel and non-lamellar structures.

Escherichia coli strain K12 was grown at 17, 27, and 37 degrees C. The acyl chain composition of the membrane lipids varied with the growth temperature; the fraction of cis-vaccenoyl chains decreased, and the fraction of palmitoyl chains increased, when the growth temperature was increased. However, the polar head group composition did not change significantly.

Influence of monoglucosyldiacylglycerol and monoacylmonoglucosyldiacylglycerol on the lipid bilayer of the membrane from Acholeplasma laidlawii strain A-EF22.

The ability for 1,2-diacyl-3-O-(alpha-D-glucopyranosyl)-sn-glycerol (MGlcDAG) and 1,2-diacyl-3-O-(6-O-acyl-(alpha-D-glucopyranosyl))-sn-glycerol (MAMGlcDAG) to induce non-lamellar phases in a lipid mixture with an in vivo composition, prepared from Acholeplasma laidlawii membranes, has been investigated. The phase transition temperatures from lamellar to non-lamellar structures were studied with varying fractions of MGlcDAG and MAMGlcDAG. The transition temperature decreased from 73 +/- 2 degrees C for 20 mol% MGlcDAG to 43 +/- 1 degree C for 63 mol% MGlcDAG, in lipid mixtures where the other lipids are the native bilayer-forming lipids.

Structure of digalactosyldiacylglycerol from oats.

Digalactosyldiacylglycerol (DGalDAG) is one of the major lipids in the cells of higher plants. DGalDAG forms a lamellar liquid crystalline phase together with water. Lipid aggregates can thus be prepared which are of potential interest for use within the cosmetic and pharmaceutical industries.

Structures of glucolipids from the membrane of Acholeplasma laidlawii strain A-EF22. III. Monoglucosyldiacylglycerol, diglucosyldiacylglycerol, and monoacyldiglucosyldiacylglycerol.

The structures of three glucolipids from the membrane of Acholeplasma laidlawii, strain A-EF22, were determined by high resolution 1H-NMR and 13C-NMR spectroscopy. The two most abundant glucolipids in this organism were shown to be 1,2-diacyl-3-O-(alpha-D-glucopyranosyl)-sn-glycerol (MGlcDAG) and 1,2-diacyl-3-O-[alpha-D-glucopyranosyl-(1 --> 2)-O-alpha-D- glucopyranosyl]-sn-glycerol (DGlcDAG). These structures agree with those determined previously by chemical analyses of the two most abundant glucolipids synthesized by the B strain of A.

Lipid extracts from membranes of Acholeplasma laidlawii A grown with different fatty acids have a nearly constant spontaneous curvature.

X-ray diffraction methods were used to explore the variation in the spontaneous curvature of lipid extracts from Acholeplasma laidlawii strain A-EF22 grown with different mixtures of palmitic acid and oleic acid. It was shown that the cells respond to the different growing conditions by altering the polar head group compositions in order to keep the phase transition between lamellar and nonlamellar structures within a narrow temperature range. This has been interpreted to mean that the membrane lipids are adjusted toward an optimal packing (Lindblom et al.

Membrane lipid composition and cell size of Acholeplasma laidlawii strain A are strongly influenced by lipid acyl chain length.

The small, cell-wall-less prokaryote Acholeplasma laidlawii strain A-EF22 could grow with membrane lipids having an average acyl chain length Cn varying over 14.5- almost 20 carbons by exogenous supplementation with selected fatty acids. For 16 < Cn < 18, the cells grew with lipids containing 100% (mol/100 mol) monounsaturated acyl chains, whereas for Cn < 16 and Cn > 18, cell growth only occurred with gradually lower fractions of unsaturated chains.

Structures of glucolipids from the membrane of Acholeplasma laidlawii strain A-EF22. II. Monoacylmonoglucosyldiacylglycerol.

The structure of one glucolipid from the membrane of Acholeplasma laidlawii, strain A-EF22, was determined. This glucolipid is synthesized only when a large fraction of saturated, straight-chain fatty acids are incorporated into the membrane lipids of strain A-EF22. The lipid was studied by 1H- and 13C-NMR spectroscopy.

Membrane thickness and molecular ordering in Acholeplasma laidlawii strain A studied by 2H NMR spectroscopy.

Since Acholeplasma laidlawii can be restricted to incorporating fatty acids from the growth medium into its membrane lipids, it is possible to study the effects of the length of the acyl chains on the properties of the membrane of the organism. A. laidlawii strain A-EF22 was grown with mixtures of one perdeuterated saturated fatty acid and one monounsaturated fatty acid.

Structures of glucolipids from the membrane of Acholeplasma laidlawii strain A-EF22. I. Glycerophosphoryldiglucosyldiacylglycerol and monoacylbisglycerophosphoryldiglucosyldiacylglycerol.

The structures of two phosphoglucolipids from the membrane of Acholeplasma laidlawii, strain A-EF22 were determined by high resolution 13C-, 31P- and 1H-NMR. The lipids in question are 1,2-diacyl-3-O-[glycerophosphoryl-6-O-(alpha-D-glucopyranosyl- (1-->2)-O-alpha-D-glucopyranosyl)]-sn-glycerol (1) and 1,2-diacyl-3-O-[glycerophosphoryl-6-O-(alpha-D-glucopyranosyl-(1-- >2)- monoacyl-glycerophosphoryl-6-O-alpha-D-glucopyranosyl)]-sn-glycero l (2). Both lipids are thus derivatives of diglucosyldiacylglycerol.

Thermotropic phase properties of 1,2-di-O-tetradecyl-3-O-(3-O-methyl- beta-D-glucopyranosyl)-sn-glycerol.

The hydration properties and the phase structure of 1,2-di-O-tetradecyl-3-O(3-O-methyl-beta-D-glucopyranosyl)-sn-glycerol (3-O-Me-beta-D-GlcDAIG) in water have been studied via differential scanning calorimetry, 1H-NMR and 2H-NMR spectroscopy, and x-ray diffraction. Results indicate that this lipid forms a crystalline (Lc) phase up to temperatures of 60-70 degrees C, where a transition through a metastable reversed hexagonal (Hll) phase to a reversed micellar solution (L2) phase occurs. Experiments were carried out at water concentrations in a range from 0 to 35 wt%, which indicate that all phases are poorly hydrated, taking up < 5 mol water/mol lipid.

Regulation and phase equilibria of membrane lipids from Bacillus megaterium and Acholeplasma laidlawii strain A containing methyl-branched acyl chains.

Phosphatidylethanolamine (PE) was isolated from Bacillus megaterium grown at 20 and 55 degrees C (PE-20 and PE-55). Iso and anteiso methyl-branched, saturated acyl chains are predominant in B. megaterium, and the value of the molar ratio of iso/anteiso acyl chains is more than 20-fold higher in PE-55 than in PE-20.