J. Lipid Res.
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Yokoyama, K.
Right arrow Articles by Setaka, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Yokoyama, K.
Right arrow Articles by Setaka, M.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Journal of Lipid Research, Vol. 41, 142-147, January 2000
Copyright © 2000 by Lipid Research, Inc.

Methods

Simultaneous separation of lysophospholipids from the total lipid fraction of crude biological samples using two-dimensional thin-layer chromatography

Kazukaki Yokoyamaa, Fuzuki Shimizua, and Morio Setakaa
a Faculty of Pharmaceutical Sciences, Teikyo University, Sagamiko, Tsukui, Kanagawa 199-0195, Japan

Correspondence to: Kazukaki Yokoyama


  ABSTRACT
TOP
 ABSTRACT
INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES

A novel solvent system for two-dimensional thin-layer chromatography was shown to simultaneously separate lysophospholipid standards, including lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylserine, lysophosphatidylinositol, lysophosphatidylglycerol, lysophosphatidic acid, lysosphingomyelin (sphingosylphosphorylcholine), and sphingosine-1-phosphate from diradylphospholipids, glycosphingolipids, and neutral lipids. Lysophospholipids contained in the total lipid fraction of activated platelets were also well separated by the same system. The present system is a useful tool for the metabolic and structural analysis of lysophospholipids in biological samples.—Yokoyama, K., F. Shimizu, and M. Setaka. Simultaneous separation of lysophospholipids from the total lipid fraction of crude biological samples using two-dimensional thin-layer chromatography. J. Lipid Res. 2000. 41: 142–147.

Supplementary key words: lysophospholipids, two-dimensional thin-layer chromatography, lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylserine, lysophosphatidylinositol, lysophosphatidylglycerol, lysophosphatidic acid, sphingosylphosphorylcholine, sphingosine-1-phosphate


  INTRODUCTION
TOP
 ABSTRACT
 INTRODUCTION
MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES

methods

Lysophospholipids (lysoPLs) have been accepted as intermediate products of the phospholipid deacylation–reacylation pathway (1) (2). Lysophosphatidic acid (lysoPA) is also synthesized de novo by esterification of glycerol-3-phosphate in the glycerolipid biosynthetic pathway (1) (3). Recently, numerous reports have revealed that lysoPLs show various biological activities themselves. For instance, lysophosphatidylcholine (lysoPC) induces intracellular calcium mobilization (4), gene expression of various proteins (5) (6) (7) (8) (9) (10) (11) (12), and is a chemotaxin for monocytes (13). LysoPA and sphingosine-1-phosphate (Sp1p) bind to very similar but specific receptors coupled to G-proteins (14) (15) (16) (17) (18) and induce various biological responses, such as cell proliferation, platelet aggregation, and intracellular stress fiber formation (14) (15) (16). In addition, Sp1P protects cells from apoptosis (16). Sphingosylphosphorylcholine (lysoSM) shares its receptor with Sp1P (19) (20) (21) and also acts as a mitogen (22), as a stimulator of smooth muscle contraction (23), as an activator of various protein kinases and an ion exchanger (20) (24) (25), presumably via intracellular calcium mobilization (19). Lysophosphatidylglycerol (lysoPG) inhibits lysoPA (26). Lysophosphatidylinositol (lysoPI) shows mitogenic activity (27). Lysophosphatidylserine (lysoPS) acts as a potent coactivator for mast cells (28). Lysophosphatidylethanolamine (lysoPE) affects the regulated exocytosis (29).

These findings emphasize the need for analysis of the production and localization of lysoPLs at their sites of function, which means that the extraction and identification of lysoPLs from crude preparations of cells and tissues is required. Only a few attempts have been made to detect the lysoPLs present in biological samples, in contrast to the many functional studies. Furthermore, the sites at which lysoPLs were detected in previous studies were not always coincident with the expected target tissues and cells. For example, lysoPCs accumulate in oxidized low density lipoprotein and atherosclerotic aorta (30) (31) and in lesions of ischemia (32). LysoPA and Sp1P are produced and released from activated platelets (33) (34). LysoPI accumulates in stimulated macrophages and oncogene-transformed fibroblasts (27) (35). LysoPS is produced in leukocytes and activated platelets (36) (37). LysoPE accumulates in serum (38) and ischemic heart (39) and is released from cultured hepatocytes (40). LysoSM is present in meningioma cells (41). LysoPG accumulates in crystalline lens (42). Moreover, various methods were used in these studies such as column chromatography (38) (39), nuclear magnetic resonance spectroscopy (41) (42), and thin-layer chromatography (TLC) with silica gel plates (27) (29) (30) (31) (34) (35) (36) (37) (40). Each of these methods was designed to separate a specific lysoPL class, which means that a standard method for separating all lysoPLs from other lipids in biological samples has not been available.

We decided to establish a simple and reproducible method for the simultaneous separation of lysoPLs from other lipids using the most widely used system, two-dimensional TLC with silica gel plates. We previously developed a method for the simultaneous separation of glycosphingolipids from other lipids in the total lipid fraction from crude biological samples using two-dimensional TLC (43) (44). Therefore, we began by modifying this system for the separation of lysoPLs. Here, we report a system that simultaneously separates lysoPLs in crude biological samples from diradylphospholipids and sphingolipids on a TLC plate.


  MATERIALS AND METHODS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
RESULTS
 DISCUSSION
 REFERENCES

Lipid standards
LysoPC (1-oleoyl-), lysoPE (1-oleoyl-), lysoPS (sodium salt, 1-oleoyl-), lysoPG (sodium salt, 1-oleoyl-), lysoPA (sodium salt, 1-oleoyl-), phosphatidylcholine (PC) (chicken egg), phosphatidylethanolamine (PE) (chicken egg), phosphatidylserine (PS) (sodium salt, 1-palmitoyl-2-oleoyl-), phosphatidic acid (PA) (sodium salt, derived from chicken egg PC), phosphatidylglycerol (PG) (sodium salt, derived from chicken egg PC), and sphingomyelin (SM) (bovine brain) were purchased from Avanti Polar Lipids, Alabaster, AL. Phosphatidylinositol (PI) (ammonium salt, soybeans), lysoPI (ammonium salt, derived from porcine pancreas PI), cholesteryl oleate, oleic acid, monoolein, and triolein were purchased from Doosan Serdary Res. Lab, Englewood Cliffs, NJ. LysoPC (1-caproyl-), lysoPC (1-decanoyl-), cardiolipin (sodium salt, bovine heart), diolein, sphingosine, and ceramide (Cer) (derived from chicken egg SM) were purchased from Sigma, St. Louis, MO. Platelet-activating factor (PAF) and Sp1P were obtained from Cayman Chemical, Ann Arbor, MI. A mixture of neutral glycosphingolipids (GlcCer, LacCer, Gb3Cer, and Gb4Cer) and a mixture of bovine brain gangliosides (GM1, GD1a, GD1b, and GT1b) were products of Iatron, Tokyo, Japan. LysoSM, lysoPAF, GM3, and free cholesterol were purchased from Biomol Research Lab, Plymouth Meeting, PA, Cascade Biochem, Berkshire, UK, Snow Brand Milk, Tokyo, Japan, and Wako, Osaka, Japan, respectively. CyclicPA (1-oleoyl-) was a gift from Dr. Tetsuyuki Kobayashi (Ochanomizu University). LysoPC (1-melissoyl-) was produced by the action of porcine pancreas phospholipase A2 from 1-melissoyl-2-oleolyl-PC derived from a mutant strain of Schizosaccharomyces pombe. Its structure was confirmed by gas chromatography and mass spectrometry (K. Yokoyama, S. Saitoh, F. Shimizu, K. Nakamura, A. Tokumura, Y. Yamakawa, M. Nishijima, M. Yanagida, and M. Setaka, unpublished results).

Two-dimensional TLC
Authentic standards were spotted in the lower left-hand corner of a TLC plate (Merck, Darmstadt, Germany, #5641, 10 cm x 10 cm). The first chromatographic run was performed with chloroform–methanol–formic acid–water 60:30:7:3 (by vol). The second run was performed with chloroform–methanol–28% ammonia in water–water 50:40:8:2 (by vol) at 90° to the original direction. When needed to separate the lysoPLs from the neutral lipids on the same plate, a third run was performed with diethylether in the opposite direction to the second run. The plate was sprayed with 20% (by vol) H2SO4 and the spots were visualized by heating.

Preparation of a biological sample containing lysoPLs
Washed rat platelets were prepared as described as previously (37) with modifications. Blood was drawn from a male Wistar rat by cardiac puncture, anticoagulated with acid citrate dextrose, and platelet-rich plasma was obtained by centrifuging at 300 g for 10 min. Platelets were washed with 137 mM NaCl, 12 mM NaHCO3, 0.4 mM Na2HPO4, 2 mM citric acid, 2.7 mM KCl, and 0.1% glucose (w/v) (pH 6.5) by centrifuging at 1,500 g for 10 min and resuspended in 10 mM Tris-HCl, 137 mM NaCl, 2.7 mM KCl, and 0.1% glucose (w/v) (pH 7.4). The washed platelets were activated with 2 µM A23187 (Calbiochem, La Jolla, CA) in the presence of 2 mM CaCl2 for 1 h and the total lipid fraction was extracted according to Bligh and Dyer (45) three times, adding citric acid to the aqueous phase at the second extraction at a final concentration of 200 µM. The chromatographic run was performed as described above.


  RESULTS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
DISCUSSION
 REFERENCES

Separation of lysoPLs standards from other lipid standards
Each lysoPL (lysoPC, lysoPE, lysoPS, lysoPI, lysoPA, lysoPG, lysoSM, Sp1P) was clearly separated from the phospholipids (PC, PE, PS, PI, PA, PG, SM, and cardiolipin), neutral glycosphingolipids (GlcCer, LacCer, Gb3Cer, and Gb4Cer) and gangliosides (GM3 and bovine brain ganglioside mixture) and were seen as distinct spots on the TLC plate ( Figure 1A). Neutral lipids, which were seen in the upper right-hand corner of the plate after the second run, could be roughly separated as larger spots in the third run. When it is not necessary to separate the neutral lipids, the third run can be omitted, because the polar lipids (including the lysoPLs) were developed after the second run and did not migrate during the third run (Figure 1B). Table 1 shows the Rf values of the lysophospholipids.



View larger version (58K):
[in this window]
[in a new window]
  		Figure 1. Separation of lysophospholipids from diradylphospholipids and sphingolipids on a two-dimensional thin-layer chromatography (TLC) plate. (A) Authentic standards were spotted on the lower left-hand corner of a TLC plate. The first chromatographic run was performed with chloroform–methanol–formic acid–water 60:30:8:2 (by vol). Then the plate was developed with chloroform–methanol–28% ammonia in water–water 50:40:7:3 (by vol) for the second dimension. The third run was performed with diethylether in the opposite direction to the second run. The plate was sprayed with 20% (by vol) H2SO4 and the spots were visualized by heating. (B) The lipids were spotted and the plate was developed as for (A) except that the third run was omitted. (C) Schematic diagram of the separation pattern. Lysophospholipids are shaded.


 
View this table:
[in this window]
[in a new window]
  		Table 1. Rf values of lysoPLs

Differences in Rf values among molecular species
The effect of fatty chain length on Rf value was examined. LysoPC with shorter fatty chains (C6:0 and C10:0) migrated to a lesser degree than lysoPC (C18:1). Conversely, a longer chain length (C30:0) resulted in greater migration (Table 1). Then, the different salt forms of acidic lysophospholipids were examined. Sodium salts of lysoPA and lysoPS were converted to the free form or calcium salts by solvent partition (46) (Table 1). Their mobilities in both directions were not affected by the salt forms, although calcium salts showed tailing to some extent (data not shown). Alkyl- and alkenyl-type lysoPC co-migrated with acyl-type lysoPC (Table 1), indicating that the linkage between glycerol and the fatty chain did not affect the Rf value, and thus these lysoPCs including lysoPAF could not be separated in this system. PAF and cyclicPA co-migrated with the lower spot of SM and PE, respectively.

Separation of lysoPLs contained in activated platelets
Figure 2B shows the TLC pattern of the total lipids of activated rat platelets. A decrease in PC, PE, and PS and a concomitant increase in lysoPC, lysoPE, lysoPS, and free fatty acid were observed compared with the total lipids of the resting platelets (Figure 2A), suggesting that platelet phospholipids were degraded by extracellularly released platelet secretory phospholipase As upon platelet activation (37). The lysoPLs even in the biological sample were clearly separated from diradylphospholipids and sphingolipids on a single plate.



View larger version (52K):
[in this window]
[in a new window]
  		Figure 2. Separation pattern of lysophospholipids contained in a biological sample. (A) Total lipid fraction of rat washed platelets. (B) The platelets were activated with 2 µM A23187 in the presence of 2 mM CaCl2 for 1 h and the total lipid fraction was extracted. The chromatographic run was performed as described in the legend to Figure 1.


  DISCUSSION
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
REFERENCES

Here, we report a simple method for the simultaneous separation of lysoPLs in biological samples from diradylphospholipids, glycosphingolipids, and neutral lipids. The lysoPLs isolated on the TLC plate can be recovered by scraping them from the plate with the silica, followed by extraction, and are available for structural analysis such as molecular mass analysis by mass spectrometry, fatty acid analysis, and the quantification of phosphorus. Alternatively, this system could be used to analyze lysoPL metabolism. A metabolic labeling study using various radiolabeled precursors of the phospholipids should be more informative. As the polar lipids do not migrate during the third run, appropriate neutral lipids, which do not incorporate radioactivity, can be used as internal standards (43). In some cases, the molecular species of the lysoPLs should be considered upon application of this system, as fatty chains of unusual length, such as 6:0 and 30:0, brought about differences in mobility. On the other hand, the linkage between glycerol and the fatty chain and the different salt forms of acidic lysoPLs did not significantly affect the Rf values. Thus, the recently identified biologically functional molecule, alkenyl-type lysoPA (46), might be developed with acyl-type lysoPA. On the other hand, calcium salts of acidic lysoPLs should be avoided by removing calcium ions by extraction under acidic and/or chelating conditions (47) (48), because they show low extraction efficiency (48) and tailing in TLC. The lysoPLs analyzed in this study were only those of the 1-acyl-2-lyso form. Application of this system to 1-lyso-2-acyl form lysoPLs will be complicated by the difference in Rf values and their conversion to the 1-acyl-2-lyso form by acyl residue migration during the treatment. Our preliminary trial to separate PAF and lysoPAF from SM and lysoPC by changing the solvent composition did not succeed, therefore sphingomyelinase-treatment is required for PAF analysis using this system.

LysoPLs with glycerol backbones are degradation products of phospholipase A2 and these lysoPLs can be metabolized by lysophospholipase and acyltransferase; therefore, their accumulation in cells and tissues indicates phospholipase A2 activation. Recently, a diverse family of phospholipase A2 isoforms has been cloned and found to tightly regulate cell signaling via production of the precursors of lipid mediators, that is, lysoPLs and free fatty acids including arachidonic acid (49). Most of them show distinct substrate specificities in vitro (50). Therefore, to better understand phospholipase A2 isoform activation, we can study lysoPL accumulation in cells and tissues. On the other hand, to determine the in vivo substrate specificity of a phospholipase A2 isoform, the class of lysoPL that accumulates in recombinant phospholipase A2-transfected cells can be examined. The present system is a useful tool for analyzing these problems.


  ACKNOWLEDGMENTS

This work was partly supported by a Grant-in-aid for Scientific Research from the Ministry of Education, Sciences, Sports and Culture of Japan. We thank Dr. Tetsuyuki Kobayashi for providing cyclicPA and for useful discussion.

Manuscript received May 24, 1999; and in revised form October 19, 1999

Abbreviations: Cer, ceramide; Gb3Cer, Gal({alpha}1-4)Gal(ß1-4)Glc(ß1-1')Cer; Gb4Cer, GalNAc(ß1-3)Gal({alpha}1-4)Gal(ß1-4)Glc(ß1-1')Cer; GD1a, NeuAc({alpha}2-3)Gal(ß1-3)GalNAc(ß1-4)[NeuAc({alpha}2-3)]Gal(ß1-4) Glc(ß1-1')Cer; GD1b, Gal(ß1-3)GalNAc(ß1-4)[NeuAc({alpha}2-8)NeuAc({alpha}2-3)] Gal (ß1-4)Glc(ß1-1')Cer; GlcCer, Glc(ß1-1')Cer; GM1, Gal(ß1-3) GalNAc (ß1-4)[NeuAc({alpha}2-3)]Gal(ß1-4)Glc(ß1-1')Cer; GM3, NeuAc({alpha}2-3)Gal(ß1-4) Glc(ß1-1')Cer; LacCer, Gal(ß1-4)Glc(ß1-1')Cer; GT1b, NeuAc({alpha}2-3)Gal(ß1-3)GalNAc(ß1-4)[NeuAc({alpha}2-8)NeuAc({alpha}2-3)]Gal (ß1-4)Glc (ß1-1')Cer; lysoPA, lysophosphatidic acid; lysoPC, lysophosphatidylcholine; lysoPE, lysophosphatidylethanolamine; lysoPG, lysophosphatidylglycerol; lysoPI, lysophosphatidylinositol; lysoPL, lysophospholipid; lysoPS, lysophosphatidylserine; lysoSM, sphingosylphosphorylcholine; PA, phosphatidic acid; PAF, platelet-activating factor; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PG, phosphatidylglycerol; PI, phosphatidylinositol; PS, phosphatidylserine; SM, sphingomyelin; Sp1p, sphingosine-1-phosphate; TLC, thin-layer chromatography


  REFERENCES
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES

  1. Vance, D. E. 1996. Glycerolipid biosynthesis in eukaryotes. In New Comprehensive Biochemistry. 31: Biochemistry of Lipids, Lipoproteins and Membranes. D.E. Vance and J.E. Vance, editors. Elsevier, Amsterdam, Netherlands. 153–181.

  2. Dircks, L. K., Sul, H. S. 1997. Mammalian mitochondrial glycerol-3-phosphate acyltransferase. Biochim. Biophys. Acta. 1348:17-26[Medline].

  3. Choy, P. C., Skrzypczak, M., Lee, D., Jay, F. T. 1997. Acyl-GPC and alkenyl/alkyl-GPC: acyl-CoA acyltransferases. Biochim. Biophys. Acta. 1348:124-133[Medline].

  4. Yu, L., Netticadan, T., Xu, Y-J., Panagia, V., Dhalla, N. S. 1998. Mechanisms of lysophosphatidylcholine-induced increase in intracellular calcium in rat cardiomyocytes. J. Pharmacol. Exp. Ther. 286:1-8[Abstract/Free Full Text].

  5. Kume, N., Cybulsky, M. I., Gimbrone, M. A., Jr. 1992. Lysophosphatidylcholine, a component of atherogenic lipoproteins, induces mononuclear leukocyte adhesion molecules in cultured human and rabbit arterial endothelial cells. J. Clin. Invest. 90:1138-1144.

  6. Kume, N., Gimbrone, M. A., Jr. 1994. Lysophosphatidylcholine transcriptionally induces growth factor gene expression in cultured human endothelial cells. J. Clin. Invest. 93:907-911.

  7. Hirata, K., Miki, N., Kuroda, Y., Sakoda, T., Kawashima, S., Yokoyama, M. 1995. Low concentration of oxidized low-density lipoprotein and lysophosphatidylcholine upregulate constitutive nitric oxide synthase mRNA expression in bovine aortic endothelial cells. Circ. Res. 76:958-962[Abstract/Free Full Text].

  8. Ochi, H., Kume, N., Nishi, E., Kita, T. 1995. Elevated levels of cAMP inhibit protein kinase C-independent mechanisms of endothelial platelet-derived growth factor-B chain and intercellular adhesion molecule-1 gene induction by lysophosphatidylcholine. Circ. Res. 77:530-535[Abstract/Free Full Text].

  9. Zembowicz, A., Tang, J., Wu, K. K. 1995. Transcriptional induction of endothelial nitric oxide synthase type III by lysophosphatidylcholine. J. Biol. Chem. 270:17006-17010[Abstract/Free Full Text].

  10. Zembowicz, A., Jones, S. L., Wu, K. K. 1995. Induction of cyclooxygenase-2 in human umbilical vein endothelial cells by lysophosphatidylcholine. J. Clin. Invest. 96:1688-1692.

  11. Sakata-Kaneko, S., Watatsuki, Y., Usui, T., Matsunaga, Y., Itoh, T., Nishi, E., Kume, N., Kita, T. 1998. Lysophosphatidylcholine upregulates CD40 ligand expression in newly activated human CD4+ T cells. FEBS Lett. 433:161-165[Medline].

  12. Liu-Wu, Y., Hurt-Camejo, E., Wiklund, O. 1998. Lysophosphatidylcholine induces the production of IL-1ß by human monocytes. Atherosclerosis. 137:351-357[Medline].

  13. Quinn, M. T., Parthasarathy, S., Steinberg, D. 1988. Lysophosphatidylcholine: a chemotactic factor for human monocytes and its potential role in atherogenesis. Proc. Natl. Acad. Sci. USA. 85:2805-2809[Abstract/Free Full Text].

  14. Moolenaar, W. H, Kranenburg, O., Postma, F. R., Zondag, G. C. M. 1997. Lysophosphatidic acid: G-protein signaling and cellular responses. Curr. Opin. Cell Biol. 9:168-173[Medline].

  15. Goetzl, E. J., An, S. 1998. Diversity of cellular receptors and functions for the lysophospholipid growth factors lysophosphatidic acid and sphingosine 1-phosphate. FASEB J. 12:1589-1598[Abstract/Free Full Text].

  16. Spiegel, S., Cuvillier, O., Edsall, L. C., Kohama, T., Menzeleev, R., Olah, Z., Olivera, A., Pirianov, G., Thomas, D. M., Tu, Z., van Brocklyn, J. R., Wang, F. 1998. Sphingosine-1-phosphate cell growth and cell death. Ann. N.Y. Acad. Sci. 845:11-18[Abstract/Free Full Text].

  17. Guo, Z., Liliom, K., Fischer, D. J., Bathurst, I. C., Tomei, L. D., Kiefer, M. C., Tigyi, G. 1996. Molecular cloning of a high-affinity receptor for the growth factor-like lipid mediator lysophosphatidic acid from Xenopus oocytes. Proc. Natl. Acad. Sci. USA. 93:14367-14372[Abstract/Free Full Text].

  18. Hecht, J. H., Weiner, J. A., Post, S. R., Chun, J. 1996. Ventricular zone gene (vzg-1) encodes a lysophosphatidic acid receptor expressed in neurogenic regions of the developing cerebral cortex. J. Cell Biol. 135:1071-1083[Abstract/Free Full Text].

  19. Okajima, F., Kondo, Y. 1995. Pertussis toxin inhibits phospholipase C activation and Ca2+ mobilization by sphingosylphosphorylcholine and galactosylsphingosine in HL60 leukemia cells. J. Biol. Chem. 270:26332-26340[Abstract/Free Full Text].

  20. Seufferlein, T., Rozengurt, E. 1995. Sphingosylphosphorylcholine activation of mitogen-activated protein kinase in Swiss 3T3 cells requires protein kinase C and a pertussis toxin-sensitive G protein. J. Biol. Chem. 270:24334-24342[Abstract/Free Full Text].

  21. Bünemann, M., Liliom, K., Brandts, B. K., Pott, L., Tseng, J-L., Desiderio, D. M., Sun, G., Miller, D., Tigyi, G. 1996. A novel membrane receptor with high affinity for lysosphingomyelin and sphingosine 1-phosphate in atrial myocytes. EMBO J. 15:5527-5534[Medline].

  22. Desai, N. N., Spiegel, S. 1991. Sphingosylphosphorylcholine is a remarkably potent mitogen for a variety of cell lines. Biochem. Biophys. Res. Commun. 181:361-366[Medline].

  23. Bitar, K. N., Yamada, H. 1995. Modulation of smooth muscle contraction by sphingosylphosphorylcholine. Am. J. Physiol. 269:G370-G377[Abstract/Free Full Text].

  24. Rodríguez-Fernández, J. L., Rozengurt, E. 1998. Bombesin, vasopressin, lysophosphatidic acid, and sphingosylphosphorylcholine induce focal adhesion kinase activation in intact Swiss 3T3 cells. J. Biol. Chem. 273:19321-19328[Abstract/Free Full Text].

  25. Törnquist, K., Woodside, M., Grinstein, S. 1997. Sphingosylphosphorylcholine activates an amiloride-insensitive Na+-H+-exchange mechanism in GH4C1 cells. Eur. J. Biochem. 248:394-400[Medline].

  26. Xu, Y., Fang, X-J., Casey, G., Mills, G. B. 1995. Lysophospholipids activate ovarian and breast cancer cells. Biochem. J. 309:933-940.

  27. Falasca, M., Iurisci, C., Carvelli, A., Sacchetti, A., Corda, D. 1998. Release of the mitogen lysophosphatidylinositol from H-ras-transformed fibroblasts: a possible mechanism of autocrine control of cell proliferation. Oncogene. 16:2357-2365[Medline].

  28. Tamori-Natori, Y., Horigome, K., Inoue, K., Nojima, S. 1986. Metabolism of lysophosphatidylserine, a potentiator of histamine release in rat mast cells. J. Biochem. (Tokyo). 100:581-590[Abstract/Free Full Text].

  29. Ikeda, Y., Fukuoka, S., Kito, M. 1997. Increase in lysophosphatidylethanolamine in the cell membrane upon the regulated exocytosis of pancreatic acinar AR42J cells. Biosci. Biotech. Biochem. 61:207-209[Medline].

  30. Steinbrecher, U. P., Parthasarathy, S., Leake, D. S., Witztum, J. L., Steinberg, D. 1984. Modification of low density lipoprotein by endothelial cells involves lipid peroxidation and degradation of low density lipoprotein phospholipids. Proc. Natl. Acad. Sci. USA. 81:3883-3887[Abstract/Free Full Text].

  31. Portman, O. W., Alexander, M. 1969. Lysophosphatidylcholine concentrations and metabolism in aortic intima plus inner media: effect of nutritionally induced atherosclerosis. J. Lipid Res. 10:158-165[Abstract].

  32. Corr, P. B., Gross, R. W., Sobel, B. E. 1984. Amphipathic metabolites and membrane dysfunction in ischemic myocardium. Circ. Res. 55:135-154[Free Full Text].

  33. Eichholtz, T., Jalink, K., Fahrenfort, I., Moolenaar, W. H. 1993. The bioactive phospholipid lysophosphatidic acid is released from activated platelets. Biochem. J. 291:677-680.

  34. Yatomi, Y., Ruan, F., Hakomiri, S., Igarashi, Y. 1995. Sphingosine-1-phosphate: a platelet-activating sphingolipid released from agonist-stimulated human platelets. Blood. 86:193-202[Abstract/Free Full Text].

  35. Zoeller, R. A., Wightman, P. D., Anderson, M. S., Raetz, C. R. H. 1987. Accumulation of lysophosphatidylinositol in RAW 264.7 macrophage tumor cells stimulated by lipid A precursors. J. Biol. Chem. 262:17212-17220[Abstract/Free Full Text].

  36. Mietto, L., Boarato, E., Toffano, G., Bruni, A. 1987. Lysophosphatidylserine-dependent interaction between rat leukocytes and mast cells. Biochim. Biophys. Acta. 930:145-153[Medline].

  37. Yokoyama, K., Kudo, I., Inoue, K. 1995. Phospholipid degradation in rat calcium ionophore-activated platelets is catalyzed mainly by two discrete secretory phospholipase As. J. Biochem. (Tokyo). 117:1280-1287[Abstract/Free Full Text].

  38. Misra, U. K. 1965. Isolation of lysophosphatidylethanolamine from human serum. Biochim. Biophys. Acta. 106:371-378[Medline].

  39. Corr, P. B., Snyder, D. W., Lee, B. I., Gross, R. W., Keim, C. R., Sobel, B. E. 1982. Pathophysiological concentrations of lysophosphatides and the slow response. Am. J. Physiol. 243:H187-H195.

  40. Robinson, B. S., Baisted, D. J., Vance, D. E. 1989. Comparison of albumin-mediated release of lysophosphatidylcholine and lysophosphatidylethanolamine from cultured rat hepatocytes. Biochem. J. 264:125-131[Medline].

  41. Seijo, L., Merchant, T. E., van der Ven, L. T. M., Minsky, B. D., Glonek, T. 1994. Meningioma phospholipid profiles measured by 31P nuclear magnetic resonance spectroscopy. Lipids. 29:359-364[Medline].

  42. Merchant, T. E., Lass, J. H., Meneses, P., Greiner, J. V., Glonek, T. 1991. Human crystalline lens phospholipid analysis with age. Invest. Ophthalmol. Vis. Sci. 32:549-555[Abstract/Free Full Text].

  43. Yokoyama, K., Nojiri, H., Suzuki, M., Setaka, M., Suzuki, A., Nojima, S. 1995. Clear differences in ceramide metabolism between glycosphingolipids and sphingomyelin in a human promyelocytic leukemia cell line HL-60 stimulated by a differentiation inducer. FEBS Lett. 368:477-480[Medline].

  44. Yokoyama, K., Suzuki, M., Kawashima, I., Karasawa, K., Nojima, S., Enomoto, T., Tai, T., Suzuki, A., Setaka, M. 1997. Changes in composition of newly synthesized sphingolipids of HeLa cells during the cell cycle: suppression of sphingomyelin and higher-glycosphingolipid synthesis and accumulation of ceramide and glucosylceramide in mitotic cells. Eur. J. Biochem. 249:450-455[Medline].

  45. Bligh, E. G., Dyer, W. J. 1959. A rapid method of total lipid extraction and purification. Can. J. Biochem. Physiol. 37:911-917.

  46. Liliom, K., Guan, Z., Tseng, J-L., Desiderio, D. M., Tigyi, G., Watsky, M. A. 1998. Growth factor-like phospholipids generated after corneal injury. Am. J. Physiol. 274:C1065-C1074[Abstract/Free Full Text].

  47. Rathbone, L. 1962. Some observations on the chromatographic behaviour of phosphatidylserine. Biochem. J. 85:461-466[Medline].

  48. Odavic, C. L. R., Sargent, E. J. 1967. The chemical nature of the products obtained by the action of cabbage-leaf phospholipase D on lysolecithin: the structure of lysolecithin. Biochem. J. 102:221-229[Medline].

  49. Murakami, M., Shimbara, S., Kambe, T., Kuwata, H., Winstead, M. V., Tischfield, J. A., Kudo, I. 1998. The functions of five distinct mammalian phospholipase A2s in regulating arachidonic acid release. J. Biol. Chem. 273:14411-14423[Abstract/Free Full Text].

  50. Mayer, R. J., Marshall, L. A. 1993. New insights on mammalian phospholipase A2(s): comparison of arachidonoyl-selective and -nonselective enzymes. FASEB. J. 7:339-348[Abstract].


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 J BiochemHome page
K. Yokoyama, M. Nakagawa, M. Satoh, S. Saitoh, N. Dohmae, A. Harada, N. Satoh, K. Karasawa, K. Takio, M. Yanagida, et al.
Expression of a Novel 90-kDa Protein, Lsd90, Involved in the Metabolism of Very Long-chain Fatty Acid-containing Phospholipids in a Mitosis-defective Fission Yeast Mutant
J. Biochem., March 1, 2008; 143(3): 369 - 375.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Biol.Home page
M. Bektas, S. G. Payne, H. Liu, S. Goparaju, S. Milstien, and S. Spiegel
A novel acylglycerol kinase that produces lysophosphatidic acid modulates cross talk with EGFR in prostate cancer cells
J. Cell Biol., June 6, 2005; 169(5): 801 - 811.
[Abstract] [Full Text] [PDF]

Home page
 J. Lipid Res.Home page
S. D. Nguyen and D.-E. Sok
Preferential inhibition of paraoxonase activity of human paraoxonase 1 by negatively charged lipids
J. Lipid Res., December 1, 2004; 45(12): 2211 - 2220.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
G. Liebisch, W. Drobnik, B. Lieser, and G. Schmitz
High-Throughput Quantification of Lysophosphatidylcholine by Electrospray Ionization Tandem Mass Spectrometry
Clin. Chem., December 1, 2002; 48(12): 2217 - 2224.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. van Tienhoven, J. Atkins, Y. Li, and P. Glynn
Human Neuropathy Target Esterase Catalyzes Hydrolysis of Membrane Lipids
J. Biol. Chem., May 31, 2002; 277(23): 20942 - 20948.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Yokoyama, K.
Right arrow Articles by Setaka, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Yokoyama, K.
Right arrow Articles by Setaka, M.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Journal of Biological Chemistry 
 Molecular and Cellular Proteomics   ASBMB Today