The action of cobra venom phospholipase A2 isoenzymes towards intact human erythrocytes

Roelofsen, B.; Trip, M. Sibenius; Verheij, Hubertus M.; Zevenbergen, J. L.

doi:10.1016/0005-2736(80)90504-0

Cited by 16 publications

(7 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It has been suggested that this effect is a consequence of the production of the split products, lysolecithin and free fatty acids, which cause an increase in lateral surface pressure in the outer monolayer. This pressure may reach a certain level above which the enzyme can no longer penetrate (Roelofsen et al, 1980). In agreement with the above monolayer studies, it appears that porcine AMPA in contrast to [Lau-Lys116] porcine and [Pal-Lys10] bovine AMPAs is able to degrade the lecithin in the intact human red cell and, most interestingly, to do so at a much higher rate and with greater efficiency than Naja naja PLA (Figure 5).…”

Section: Discussionsupporting

confidence: 76%

Site-specific .epsilon.-amino monoacylation of pancreatic phospholipase A2. 2. Transformation of soluble phospholipase A2 into a highly penetrating "membrane-bound" form

Wiele¹,

Atsma²,

Roelofsen³

et al. 1988

Biochemistry

View full text Add to dashboard Cite

Long-chain lecithins present in bilayer structures like vesicles or membranes are only very poor substrates for pancreatic phospholipases A2. This is probably due to the fact that pancreatic phospholipases A2 cannot penetrate into the densely packed bilayer structures. To improve the weak penetrating properties of pancreatic phospholipases A2, we prepared and characterized a number of pancreatic phospholipase A2 mutants that have various long acyl chains linked covalently to Lys116 in porcine and to Lys10 in bovine phospholipase A2 [Van der Wiele, F.C., Atsma, W., Dijkman, R., Schreurs, A.M.M., Slotboom, A.J., & De Haas, G.H. (1988) Biochemistry (preceding paper in this issue)]. When monomolecular surface layers of L- and D-didecanoyllecithin were used, it was found that the introduction of caprinic, lauric, palmitic, and oleic acid at Lys116 in the porcine enzyme increases its penetrating power from 13 to about 17, 20, 32, and 22 dyn/cm, respectively, before long lag periods were obtained. Incorporation of a palmitoyl moiety at Lys10 in the bovine enzyme shifted the penetrating power from 11 to about 25 dyn/cm. Only the best penetrating mutant, viz., porcine phospholipase A2 having a palmitoyl moiety at Lys116, was able to cause complete leakage of 6-carboxyfluorescein entrapped in small unilamellar vesicles of egg lecithin under nonhydrolytic conditions. Similarly, only this latter palmitoylphospholipase A2 completely hydrolyzed all lecithin in the outer monolayer of the human erythrocyte at a rate much faster than Naja naja phospholipase A2, the most powerful penetrating snake venom enzyme presently known.

show abstract

Section: Discussionsupporting

confidence: 76%

Site-specific .epsilon.-amino monoacylation of pancreatic phospholipase A2. 2. Transformation of soluble phospholipase A2 into a highly penetrating "membrane-bound" form

Wiele¹,

Atsma²,

Roelofsen³

et al. 1988

Biochemistry

View full text Add to dashboard Cite

show abstract

“…Phospholipases have been used extensively to determine membrane phospholipid organization in RBC from normal individuals (20)(21)(22)(23). Among these phospholipases are phospholipase A2 from bee venom and sphingomyelinase C from Staphylococcus aureus.…”

Section: Introductionmentioning

confidence: 99%

Abnormalities in membrane phospholipid organization in sickled erythrocytes.

Lubin¹,

Chiu²,

Bastacky³

et al. 1981

J. Clin. Invest.

250

115

View full text Add to dashboard Cite

A B S T R A C T In contrast to the wealth of infonnation concerning membrane phospholipid asymmetry in normal human erythrocytes, very little is known about membrane phospholipid organization in pathologic erythrocytes. Since the spectrin-actin lattice, which has been suggested to play an important role in stabilizing membrane phospholipid asymmetry, is abnormal in sickled erythrocytes, we determined the effects of sickling on membrane phospholipid organization. We used two enzymatic probes: bee venom phospholipase A2 and Staphylococcus aureus sphingomyelinase C, which do not penetrate the membrane and react only with phospholipids located in the outer leaflet of the bilayer. Our results suggest that the distribution of glycerophospholipids within the membrane of sickled cells is* different from that in nonsickled cells. Compared with the normal erythrocyte, the outer membrane leaflet of the deoxygenated, reversibly sickled cells (RSC) and irreversibly sickled cells (ISC) was enriched in phosphatidyl ethanolamine in addition to containing phosphatidyl serine. These changes were compensated for by a decrease in phosphatidyl choline in that layer. The distribution of sphingomyelin over the two halves of the bilayer was unaffected by sickling. In contrast to ISC, where the organization of phospholipids was abnormal under both oxy and deoxy conditions, reoxygenation of RSC almost completely restored the organization of membrane phospholipids to normal. These results indicate that the process of sickling induces an abnormality in the organization of membrane lipids in RSC which becomes permanent in ISC.

show abstract

“…The failure of phospholipase A 2 (either from snake or bee venom) to degrade all of its substrates present in the outer membrane layer of an erythrocyte has been well documented for the human red cell [3,23]. It has been argued [37] that the action of the phospholipase A 2 ceases as a consequence of an increase in lateral surface pressure in the outer monolayer of the intact cell, which is caused by the generation of split products (lysophospholipids and free fatty acids) herein. Hydrolysis of sphingomyelin by sphingomyelinase C is believed to have the opposite effect, namely, a decrease in lateral surface pressure [3,37].…”

Section: Discussionmentioning

confidence: 99%

“…It has been argued [37] that the action of the phospholipase A 2 ceases as a consequence of an increase in lateral surface pressure in the outer monolayer of the intact cell, which is caused by the generation of split products (lysophospholipids and free fatty acids) herein. Hydrolysis of sphingomyelin by sphingomyelinase C is believed to have the opposite effect, namely, a decrease in lateral surface pressure [3,37]. Including this enzyme in incubations with phospholipase A 2, therefore, enables the latter phospholipase to degrade its substrates in the outer monolayer of intact human erythrocytes to completion [3,23].…”

Section: Discussionmentioning

confidence: 99%

Phospholipid asymmetry during erythropoiesis. A study on Friend erythroleukemic cells and mouse reticulocytes

Schaft

Roelofsen

Kamp

et al. 1987

Biochimica et Biophysica Acta (BBA) - Biomembranes

Self Cite

View full text Add to dashboard Cite

The distribution of phospholipids over the outer and inner layers of the plasma membranes of differentiated Friend erythroleukemic cells (Friend cells) and mouse reticulocytes has been determined. Phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol were found to be distributed symmetrically over both layers, sphingomyelin was found to be enriched in the outer layer (80-85%) and phosphatidylserine appeared to be present mainly in the inner layer (80-90%) of the plasma membranes of differentiated Friend cells. The outer layer of reticulocyte membranes contains 50-60% of the phosphatidylcholine, 20% of the phosphatidylethanolamine, 82-85% of the sphingomyelin and 40-42% of the phosphatidylinositol. All of the phosphatidylserine is present in the inner layer. The results show, that the asymmetric distribution of phospholipids, typical for erythrocyte membranes, is partially apparent already at an early stage of erythropoiesis, the proerythroblast, while the final organization of phospholipid distribution takes place at some stage during enucleation of the enormoblast and release of the reticulocyte into the blood stream.

show abstract

The action of cobra venom phospholipase A2 isoenzymes towards intact human erythrocytes

Cited by 16 publications

References 22 publications

Site-specific .epsilon.-amino monoacylation of pancreatic phospholipase A2. 2. Transformation of soluble phospholipase A2 into a highly penetrating "membrane-bound" form

Site-specific .epsilon.-amino monoacylation of pancreatic phospholipase A2. 2. Transformation of soluble phospholipase A2 into a highly penetrating "membrane-bound" form

Abnormalities in membrane phospholipid organization in sickled erythrocytes.

Phospholipid asymmetry during erythropoiesis. A study on Friend erythroleukemic cells and mouse reticulocytes

Contact Info

Product

Resources

About