Biomedical subjects

T Thuren

Publications and source records attributed to T Thuren.

26 records · Page 2Linked to original sources

PubMed1987

Control of the action of phospholipases A by "vertical compression" of the substrate monolayer.

Monolayers of rac-1,2-didodecanoyl-sn-glycero-3-phosphoglycerol at an air-water interface were "vertically compressed" by substituting an alkylated glass plate for air while maintaining a constant surface pressure of 15 mN m-1. At this surface pressure the overlaying of the lipid film by the alkylated surface resulted in an average increase of 16 A2/molecule in the mean molecular area of those phospholipid molecules residing at the interface between water and the alkylated glass. Subsequently, the activities of phospholipases A1 and A2 toward the monolayers were measured both in the presence and in the absence of the support. While phospholipase A1 activity was increased 4-fold by the support, the activity of phospholipase A2 was reduced to 15% of the activity measured in the absence of the alkylated surface. These findings indicate that such a "vertical compression" of the monolayer is likely to induce a conformational change in the phospholipid molecules, which in turn would cause the above reciprocal changes in the activities of phospholipases A1 and A2. A molecular model accounting to these findings is presented.

Animals↗

PubMed1987

Triggering of the activity of phospholipase A2 by an electric field.

In this paper we show that the action of phospholipase A2 can be triggered by applying an electric field across a 1,2-didodecanoyl-sn-3-phosphoglycerol monolayer residing between an alkylated silicon surface and water. When the silicon wafer served as a cathode, rapid activation of porcine pancreatic phospholipase was observed and did depend on the magnitude of the applied potential. The degree of activation was different for the pancreatic phospholipase A2 and snake and bee venom enzymes. Maximally, a 7-fold activation of pancreatic phospholipase A2 was observed when the applied potential was 75 V. The effective field over the lipid film could be estimated to be approximately 25-175 mV, i.e., in the range of membrane potentials found in cells. On the basis of these results, we suggest that changes in membrane potential might be an important factor in the regulation of the action of intracellular phospholipases A2 in vivo.

Animals↗

PubMed1987

Hydrolysis of 1-palmitoyl-2-[6-(pyren-1-yl)]hexanoyl-sn-glycero- 3-phospholipids by phospholipase A2: effect of the polar head-group.

The effect of the phospholipid polar head-group on the porcine pancreatic phospholipase A2 (phosphatidylcholine 2-acylhydrolase, EC 3.1.1.4) reaction was studied using 1-palmitoyl-2-[6-(pyren-1-yl)]hexanoyl-sn-glycero-3- phosphatidylcholine, -ethanolamine, -glycerol, -monomethylester and -serine as substrates. Except for the monomethylester analogue, which was maximally activated by 3.5 mM CaCl2, maximal enhancement of hydrolysis of the other pyrenephospholipids was obtained at 2 mM Ca2+. Sodium cholate inhibited hydrolysis of the ethanolamine and serine lipids, whereas a slight (1.4-2.0-fold) activation was observed for the -choline, -glycerol and -monomethylester derivatives. Arrhenius plots of hydrolysis of pyrenephospholipids by porcine pancreatic phospholipase A2 revealed no discontinuities, thus indicating the absence of phase transition for these lipids in the temperature range 15-45 degrees C. Specific activities of porcine and bovine pancreatic, porcine intestinal and snake venom (Crotalus atrox) phospholipases A2 towards pyrenephospholipid liposomes were then compared. Whereas the snake venom phospholipase A2 preferred phosphatidylcholine as a substrate, the other phospholipases A2 preferred acidic phospholipids in the order monomethylester greater than or equal to glycerol greater than or equal to serine.

Animals↗

PubMed1987

Increased plasma phospholipase-A2 activity in schizophrenic patients: reduction after neuroleptic therapy.

Phospholipase-A2 (PLA2) is a key enzyme in the metabolism of phospholipids, and it may play an important role in neuronal function and neuronal plasticity. We determined the activity of PLA2 in the plasma of 20 drug-free schizophrenic patients, 6 nonschizophrenic psychiatric patients, and 21 healthy controls. Schizophrenic patients showed significantly higher plasma PLA2 activity than controls, and higher than our nonschizophrenic patients. Seventy percent of the schizophrenics had enzyme activity higher than the highest value from the control group. The increased plasma PLA2 activity in schizophrenics was reduced to the level of the controls after 3 weeks of neuroleptic treatment. These findings warrant further study for possible implications of this increased PLA2 activity in the etiopathology of schizophrenia.

Adult↗

PubMed1987

Pancreatic phospholipase A2 (PPLA2) during human pregnancy.

Pancreatic phospholipase A2 in human serum during pregnancy is within the normal range. In the amniotic fluid, the amount of immunoreactive pancreatic phospholipase A2 increases from 1.3 microgram/l at 16 weeks to 2.9 micrograms/l near term, but does not correlate with PLA2-activity. Meconium-stained amniotic fluid had highly elevated pancreatic, but not kinetic, PLA2. The fetal pancreas and its high PLA2-content may contribute to the progressive increase in uterine activity during human pregnancy.

Amniotic Fluid↗

PubMed1986

Polyamine-phospholipid interaction probed by the accessibility of the phospholipid sn-2 ester bond to the action of phospholipase A2.

Conditions were used where the action of porcine pancreatic phospholipase A2 on phospholipids can be followed in the absence of added calcium and the catalytic activity is supported by the calcium brought with the nanomolar enzyme. Therefore, alterations in the enzyme velocity resulting from the presence of spermine or spermidine could be specifically studied using 1-palmitoyl-2-(pyren-1-yl)hexanoyl-sn-glycero-3-phosphocholine (PPHPC) and 1-palmitoyl-2-(pyren-1-yl)hexanoyl-sn-glycero-3-phosphoglycerol (PPHPG) as substrates. Both spermine and spermidine activated the hydrolysis of PPHPG fourfold at polyamine/phospholipid molar ratios of approximately 1:1 and 12:1, respectively. Double-reciprocal plots of enzyme activity vs. PPHPG concentration revealed the enhancement to be due to increased apparent Vmax while the apparent Km was slightly increased. In the presence of 4 mM CaCl2 inhibition by polyamines of PPHPG hydrolysis by phospholipase A2 was observed. Using synthetic diamines we could further demonstrate that two primary amino groups are required for the activation. In the absence of exogenous CaCl2 polyamines inhibited the hydrolysis of PPHPC by phospholipase A2. The presence of 4 mM CaCl2 reversed this inhibition and a twofold activation was observed at 10 microM spermine. The results obtained indicate that the activation of PLA2 by spermine and spermidine is produced at the level of the substrate, PPHPG. This implies the formation of complexes of phosphatidylglycerol and polyamines with defined stoichiometries.

Animals↗

PubMed1985

Fluorometric assay for phospholipase A2 in serum.

In this fluorometric assay for phospholipase A2 (EC 3.1.1.4) in serum we use the fluorescent phospholipid analog 1-octacosanyl-2-(pyren-1-yl)hexanoyl-sn-glycero-3-phosphatid yl monomethyl ester as substrate. The optimized conditions are: 28 mumol of the substrate per liter of Tris buffer (20 mmol/L, pH 7.4). The hydrolytic reaction is allowed to proceed for 30 min at 37 degrees C. The fluorescent reaction product, (pyren-1-yl)hexanoic acid, is then separated from the unreacted substrate by liquid-liquid phase partition. The concentration of the liberated fatty acid analog is determined fluorometrically. The detection limit is approximately 6 pmol min-1 mL-1. Forty duplicate samples can be assayed in about 2 h. The activity of this enzyme in serum of 20 healthy volunteers averaged 69 pmol min-1 mL-1. For seven patients with clinically diagnosed pancreatitis the average activity was 1092 pmol min-1 mL-1.

Acute Disease↗

PubMedDate not supplied

Estimation of the equilibrium lateral pressure in 1-palmitoyl-2-[6(pyren-1-yl)]hexanoyl-glycerophospholipid liposomes.

Compression isotherms for 1-palmitoyl-2-[6(pyren-1-yl)] hexanoyl-sn-glycero-3-phosphocholine (PPHPC), -ethanolamine (PPHPE), -glycerol (PPHPG), -serine (PPHPS) and -phosphatidic acid monomethylester (PPHPM) were recorded at an argon/water interface. Thereafter, the ratios of pyrene excimer to monomer fluorescence emission intensities (Ie/Im) were determined for liposomes of these lipids and were found to be 20.15, 12.30, 11.80, 10.15 and 6.95 for the ethanolamine, choline, monomethylester, glycerol and serine derivatives, respectively. Assuming Ie/Im to depend on the reciprocal of the mean molecular area of the pyrenelipids in liposomes, equilibrium surface pressure to be the same in liposomes of these lipids regardless of the head group structure and neglecting any possible influence due to differences in the orientation of the pyrene moiety, we sought for that surface pressure value in the compression isotherms where the correlation of the reciprocal of mean molecular area in monolayers to Ie/Im values observed in liposomes was maximal. This treatment results in a value of approximately 12 mN m-1 for the equilibrium surface pressure in 1-palmitoyl-2-[6(pyren-1-yl)]hexanoyl-glycerophospholipid liposomes.

Liposomes↗