Lipid Biosynthesis in Peroxisomes a

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/74665/1/j.1749-6632.1996.tb18613.x.pd

Quantification, characterization and fatty acid composition of lysophosphatidic acid in different rat tissues

The amount and composition of lysophosphatidate present in different rat tissues have been estimated by an internal standard method in which a synthetic unnatural isomer (1‐heptadecanoyl‐rac‐glycerol‐3‐phosphate) was added to the total lipid extracts, and the fatty acid composition of purified lysophosphatidate was determined. Lipids from tissues were extracted under acidic conditions, and the lysophosphatidate was purified by solvent partitions followed by thin‐layer chromatography in multiple solvent systems. The purified lipid was shown to be 1‐acyl‐sn‐glycerol‐3‐phosphate by chromatographic and chemical analysis, by its resistance to hydrolysis when treated with phospholipase A2 and also by its complete conversion to 1‐acyl‐sn‐glycerol when treated with alkaline phosphatase. The fatty acid consituents of this lipid were determined by gas‐liquid chromatography of the derived methyl esters. The concentrations (nmol/g of tissue) of lysophosphatidate in various tissues were: 86.2±4.2 in brain, 60.3±6.3 in liver, 46.4±6.5 in kidney, 30.6±5.0 in testis, 22.3 in heart and 19.3 in lung. Mostly (80%) saturated fatty acids were found to be present in this lyso lipid. A significantly high level of stearic acid was present in this lipid from all the tissues (50–60% in liver, kidney, brain and testis, and about 40% in heart and lung) compared to plamitic acid (10–15% in liver, kidney and brain and 25–30% in testis, heart and lung). The fatty acid compositions of phosphatidic acid, the putative product of lysophosphatidate acylation, from different tissues were also determined and palmitate was found to be the major saturated fatty acid. These results suggest that tissue lysophosphatidic acid is not only formed byde novo biosynthesis but is also generated via the breakdown of phospholipids such as phosphoinositides.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/141048/1/lipd0329.pd

Molecular Species of Diacylglycerols and Phosphoglycerides and the Postmortem Changes in the Molecular Species of Diacylglycerols in Rat Brains

The molecular species of 1,2-diacyl- sn -glycerol (DAG), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidylinositol 4-phosphate (PIP), and phosphatidylinositol 4,5-bisphosphate (PIP 2 ) from brains of adult rats (weighing 150 g) were determined. The DAG, isolated from brain lipid extracts by TLC, was benzoylated, and the molecular species of the purified benzoylated derivatives were separated from each other by reverse-phase HPLC. The total amount and the concentration of each species were quantified by using 1,2-distearoyl- sn -glycerol (18:0–18:0) as an internal standard. About 30 different molecular species containing different fatty acids at the sn -1 and sn -2 positions of DAG were identified in rat brains (1 min postmortem), and the predominant ones were 18:0–20:4 (35%), 16:0–18:1 (15%), 16:0–16:0 (9%), and 16:0–20:4 (8%). The molecular species of PC, PE, PS, and PI were determined by hydrolyzing the lipids with phospholipase C to DAG, which was then benzoylated and subjected to reverse-phase HPLC, PIP and PIP 2 were first dephosphorylated to PI with alkaline phosphatase before hydrolysis by phospholipase C. The molecular species composition of phosphoinositides showed predominantly the 18:0–20:4 species (50% in PI and ∼65% in PIP and PIP 2 ). PS contained mainly the 18:0–22:6 (42%) and 18:0–18:1 (24%) species. PE was mainly composed of the 18:0–20:4 (22%), 18:0–22:6 (18%), 16:0–18:1 (15%), and 18:0–18:1 (15%) species. In PC the main molecular species were 16:0–18:1 (36%), 16:0–16:0 (19%), and 18:0–18:1 (14%). Studies on postmortem brains (30 s to 30 min) showed a rapid increase in the total amount (from 40–50 nmol/g in 0 min to 210–290 nmol/g in 30 min) and in all the molecular species of DAG. Comparatively larger increases (seven- to 10-fold) were found for the 18:0–20:4 and 16:0–20:4 species. Comparison of DAG species with the molecular species of different glycerolipids indicated that the rapid postmortem increase in content of DAG was mainly due to the breakdown of phosphoinositides. However, a slow but continuous breakdown of PC to DAG was also observed.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66196/1/j.1471-4159.1991.tb08161.x.pd

BIOSYNTHESIS OF PHOSPHATIDIC ACID IN RAT BRAIN VIA ACYL DIHYDROXYACETONE PHOSPHATE

The enzymes for the biosynthesis of phosphatidic acid from acyl dihydroxyacetone phosphate were shown to be present in rat brain. These enzymes were mainly localized in the microsomal fraction of 12–14 day old rat brains. The brain microsomal acyl CoA: dihydroxyacetone phosphate acyl transferase (EC 2.3.1.42), exhibited a broad pH optimum between pH 5 and 9 with maximum activity at pH 5.4. K m for DHAP at pH 5.4 was 0.1 mm and V max was 0.86nmol/min/mg of microsomal protein. The corresponding microsomal enzyme for the glycerophosphate pathway (acyl CoA: sn -glycerol-3-phosphate acyl transferase EC 2.3.1.15) was shown to have a different pH optimum (pH 7.6). On the basis of the differences in pH optima, differential effects of sodium cholate in the enzymes and a common substrate competition study, these acyl transferases were postulated to be two different microsomal enzymes. Acyl DHAP:NADPH oxidoreductase (EC 1.1.1.101) in brain microsomes was found to be quite specific for NADPH as cofactor, being able to utilize NADH only at very high concentrations. This enzyme exhibited a K m of 8.6 Μm with NADPH and V mx of 0.81 nmol/min/mg protein. The presence of these two enzymes and the known presence of l-acyl- sn -glycerol-3-phosphate: acyl CoA acyl transferase in brain (Fleming & Hajra, 1977) demonstrated the biosynthesis of phosphatidic acid in brain via acyl dihydroxyacetone phosphate. Phosphatidic acid was shown to form when dihydroxyacetone phosphate, acyl CoA, NADPH and other cofactors were incubated together with brain microsomes. Further properties of the enzymes and the probable importance of the presence of this pathway in brain were discussed.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66377/1/j.1471-4159.1978.tb12440.x.pd

Measurement of Gibbs energies of formation of CoF₂ and MnF₂ using a new composite dispersed solid electrolyte

Gibbs energies of formation of CoF2 and MnF2 have been measured in the temperature range from 700 to 1100 K using Al2O3-dispersed CaF2 solid electrolyte and Ni+NiF2 as the reference electrode. The dispersed solid electrolyte has higher conductivity than pure CaF2 thus permitting accurate measurements at lower temperatures. However, to prevent reaction between Al2O3 in the solid electrolyte and NiF2 (or CoF2) at the electrode, the dispersed solid electrolyte was coated with pure CaF2, thus creating a composite structure. The free energies of formation of CoF2 and MnF2 are (± 1700) J mol−1; ΔG°f(CoF2,s)=-664,010+134.38T(700-980K) ΔG°f(MnF2,s)=-848.270+129.59T(700-980K) =-850.200+131.56T(980-1100K) The third law analysis gives the enthalpy of formation of solid ΔG°f2 as ΔH° (298·15 K) = −672·69 (± 0·1) kJ mol−1, which compares with a value of −671·5 (± 4) kJ mol−1 given in Janaf tables. For solid MnF2, ΔH°(298·15 K) = − 854·97 (± 0·13) kJ mol−1, which is significantly different from a value of −803·3 kJ mol−1 given in the compilation by Barinet al

Biosynthesis of Triglyceride and Other Fatty Acyl Esters by Developing Rat Brain

The biosynthesis of triglyceride from 1,2-diglyceride and long-chain acyl coenzyme A (CoA) was studied in developing rat brain. Diglyceride acyltransferase activity was highest in a microsomal fraction, had a neutral pH optimum, and was stimulated by MgCl 2 . Palmitoyl CoA and oleoyl CoA served equally well as acyl donors. The enzyme catalyzed the acylation of both endogenous diglyceride and several naturally occurring and synthetic exogenous diglycerides. In addition, short-chain primary and secondary alcohols were found to be acylated under these conditions. A second acylation system, active at low pH, was found to catalyze esterification of ethanol and cholesterol, but not diglyceride, with free fatty acid. These results demonstrate that brain has the capacity to acylate a wide variety of physiological and nonphysiological hydroxyl compounds.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66158/1/j.1471-4159.1984.tb12842.x.pd

Design of a photosystem to harvest visible-light into electrons: photosensitised one electron redox reactions in organic synthesis

Based on synchronous oxidation-reduction processes, analogous to photosynthetic mechanistic paradigm, a photosystem utilising Ph3P or ascorbic acid as sacrificial electron donor has been developed to halvest electrons from visible light photons. The utility of such photosystem has been demonstrated by initiating various one-electron reductive -C-C-bo nd formation reactions. Biologically active PGE1, and C-Furanosides are synthesised employing this photosystem at a crucial step

TOPOGRAPHICAL DISTRIBUTION OF LIPID BIOSYNTHETIC ENZYMES ON PEROXISOMES (MICROBODIES)

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/72338/1/j.1749-6632.1982.tb21437.x.pd

Electrical transport in magnesium aluminate

The conductivity of MgAl2O4 has been measured at 1273, 1473 and 1673 K as a function of the partial pressure of oxygen ranging from 105 to 10−14 Pa. The MgAl2O4 pellet, sandwiched between two platinum electrodes, was equilibrated with a flowing stream of either Ar + O2, CO + CO2 or Ar + H2 + H2O mixture of known composition. The gas mixture established a known oxygen partial pressure. All measurements were made at a frequency of 1 kHz. These measurements indicate pressure independent ionic conductivity in the range 1 to 10−14 Pa at 1273 K, 10−1 to 10−12 Pa at 1473 K and 10−1 to 10−4 Pa at 1673 K. The activation energy for ionic conduction is 1·48 eV, close to that for self-diffusion of Mg2+ ion in MgAl2O4 calculated from the theoretical relation of Glyde. Using the model, the energy for cation vacancy formation and activation energy for migration are estimated