Stereochemistry- and Concentration-Dependent Effects of Phosphatidylserine Enrichment on Platelet Function (original) (raw)
Related papers
Analytical Characterization of the Role of Phospholipids in Platelet Adhesion and Secretion
Analytical Chemistry, 2015
The cellular phospholipid membrane plays an important role in cell function and cell−cell communication, but its biocomplexity and dynamic nature presents a challenge for examining cellular uptake of phospholipids and the resultant effects on cell function. Platelets, small anuclear circulating cell bodies that influence a wide variety of physiological functions through their dynamic secretory and adhesion behavior, present an ideal platform for exploring the effects of exogenous phospholipids on membrane phospholipid content and cell function. In this work, a broad range of platelet functions are quantitatively assessed by leveraging a variety of analytical chemistry techniques, including ultraperformance liquid chromatography−tandem electrospray ionization mass spectrometry (UPLC−MS/MS), vasculature-mimicking microfluidic analysis, and single cell carbon-fiber microelectrode amperometry (CFMA). The relative enrichments of phosphatidylserine (PS) and phosphatidylethanolamine (PE) were characterized with UPLC−MS/MS, and the effects of the enrichment of these two phospholipids on both platelet secretory behavior and adhesion were examined. Results show that, in fact, both PS and PE influence platelet adhesion and secretion. PS was enriched dramatically and decreased platelet adhesion as well as secretion from δ-, α-, and lysosomal granules. PE enrichment was moderate and increased secretion from platelet lysosomes. These insights illuminate the critical connection between membrane phospholipid character and platelet behavior, and both the methods and results presented herein are likely translatable to other mammalian cell systems.
Biochemical Journal, 1985
Platelets rapidly convert 1,2-didecanoyl-sn-glycerol into itscorresponding phosphatidic acid and lysophosphatidic acid derivatives, thereby providing a means of introducing these two compounds into platelets. l-Decanoyl-2-lyso-3-sn-phosphatidic acid, when added directly to platelets, induced platelet aggregation and raised intracellular Ca2+ levels at concentrations of 0.3,M upwards, but was without effect when formed intracellularly from 1,2-didecanoylglycerol at an estimated concentration of approx. 47 /SM. This indicates that the site of platelet activation by lysophosphatidic acid is extracellular. A concentration of thrombin (0.2 unit/ml), which produced maximal platelet aggregation, caused an estimated intracellular formation of 20 /M-lysophosphatidic acid in the presence of 2 mM-Ca2+; however, there was no detectable release of lysophosphatidic acid into the bathing medium. Lysophosphatidic acid, therefore, may not be an intracellular second messenger involved in platelet aggregation by thrombin.
Exposure of phosphatidylserine on the cell surface
Cell Death & Differentiation, 2016
Phosphatidylserine (PtdSer) is a phospholipid that is abundant in eukaryotic plasma membranes. An ATP-dependent enzyme called flippase normally keeps PtdSer inside the cell, but PtdSer is exposed by the action of scramblase on the cell's surface in biological processes such as apoptosis and platelet activation. Once exposed to the cell surface, PtdSer acts as an 'eat me' signal on dead cells, and creates a scaffold for blood-clotting factors on activated platelets. The molecular identities of the flippase and scramblase that work at plasma membranes have long eluded researchers. Indeed, their identity as well as the mechanism of the PtdSer exposure to the cell surface has only recently been revealed. Here, we describe how PtdSer is exposed in apoptotic cells and in activated platelets, and discuss PtdSer exposure in other biological processes.
The Ukrainian Biochemical Journal, 2017
Plasminogen/plasmin system is known for its ability to support hemostatic balance of blood. however, plasminogen may be considered as an adhesive ligand and in this way could affect the functioning of blood cells. We showed that exogenous Lys-plasminogen, but not its Glu-form, inhibited platelet aggregation and suppressed platelet α-granule secretion. The aim of this work was to investigate the influence of Glu-and Lysform of plasminogen on the formation of platelet procoagulant surface using phosphatidylserine exposure as a marker. human platelets were obtained from human platelet-rich plasma (donors were healthy volunteers, men aged 30-40 years) by gel-filtration on Sepharose 2B. Phosphatidylserine exposure on the platelet surface was evaluated by flow cytometry with FITC-conjugated annexin A5. Glu-and Lys-plasminogen have different impact on the platelet functioning. Exogenous Lys-plasminogen has no significant effect on phosphatidylserine exposure, while Glu-plasminogen increases phosphatidylserine exposure on the surface of thrombin-and collagen-activated human platelets. Glu-plasminogen can be considered as a co-stimulator of agonist-induced platelet secretion and procoagulant surface formation. Meanwhile effects of Lys-plasminogen are probably directed at platelet-platelet interactions and not related to agonist-stimulated pro-apoptotic changes. The observed different effects of Glu-and Lys-plasminogen on phosphatidylserine exposure can be explained by their structural peculiarities. k e y w o r d s: Glu-and Lys-plasminogen, phosphatidylserine exposure, platelets.
PLoS ONE, 2013
Recently, by employing intra-vital confocal microscopy, we demonstrated that platelets expose phosphatidylserine (PS) and fibrin accumulate only in the center of the thrombus but not in its periphery. To address the question how exposure of platelet anionic phospholipids is regulated within the thrombus, an in-vitro experiment using diluted platelet-rich plasma was employed, in which the fibrin network was formed in the presence of platelets, and PS exposure on the platelet surface was analyzed using Confocal Laser Scanning Microscopy. Almost all platelets exposed PS after treatment with tissue factor, thrombin or ionomycin. Argatroban abrogated fibrin network formation in all samples, however, platelet PS exposure was inhibited only in tissue factor-and thrombin-treated samples but not in ionomycin-treated samples. FK633, an a IIb b 3 antagonist, and cytochalasin B impaired platelet binding to the fibrin scaffold and significantly reduced PS exposure evoked by thrombin. Gly-Pro-Arg-Pro amide abrogated not only fibrin network formation, but also PS exposure on platelets without suppressing platelet binding to fibrin/fibrinogen. These results suggest that outside-in signals in platelets generated by their binding to the rigid fibrin network are essential for PS exposure after thrombin treatment.
Biochimica et Biophysica Acta (BBA) - Biomembranes, 1986
Human platelets in three physiological states were prepared. These states were the gel-filtered, the thrombin-induced shape-changed, and the thrombin-activated platelets. The phospholipid distributions in these three types of membrane were probed by using the basic phospholipase A 2 of Naja nigricollis. This enzyme could penetrate through these membranes to hydrolyze all of their accessible phospholipids and to cause cell lysis. The hydrolytic time-courses displayed three phases. The state of platelet in each lipid hydrolytic phase was examined by: (1) measuring the leakage of lactate dehydrogenase; (2) analyzing the morphology by both scanning and transmission electron microscopy (scanning EM and transmission EM); and (3) estimating the hydrolysis of the [32P]phosphate-labeled platelets. The existence of these three hydrolytic phases may signify that the phospholipase A 2 sequentially hydrolyzed its substrates in the membrane outer leaflet, in the inner one, and in the cytosol. The content and the distribution of each phospholipid class in the plasma membranes of the resting and of the shape-changed platelets were similar. These membrane surfaces consisted mainly of phosphatidylcholine (PC) and phosphatidylethanolamine (PE). Phosphatidylserine (PS) was not exposed on the surface of the shape-changed platelet. The content of each lipid class in the activated platelet membrane was 10% more than that in the resting platelet. PS was found on the activated platelet cell surface. This implies that PS is exposed only during platelet secretion.