Advertisement
JBC

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Originally published In Press as doi:10.1074/jbc.C300563200 on January 19, 2004

J. Biol. Chem., Vol. 279, Issue 12, 10833-10836, March 19, 2004
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Supplemental Data
Right arrow All Versions of this Article:
279/12/10833    most recent
C300563200v1
Right arrow Submit a Letter to Editor
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
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 arrowRequest Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by van Meeteren, L. A.
Right arrow Articles by Moolenaar, W. H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by van Meeteren, L. A.
Right arrow Articles by Moolenaar, W. H.
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?

ACCELERATED PUBLICATIONS

Spider and Bacterial Sphingomyelinases D Target Cellular Lysophosphatidic Acid Receptors by Hydrolyzing Lysophosphatidylcholine*

Laurens A. van Meeteren, Floor Frederiks, Ben N. G. Giepmans, Matheus F. Fernandes Pedrosa{ddagger}, Stephen J. Billington§, B. Helen Jost§, Denise V. Tambourgi{ddagger}, and Wouter H. Moolenaar

From the Division of Cellular Biochemistry and Centre for Biomedical Genetics, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands, the {ddagger}Laboratório de Imunoquímica, Instituto Butantan, 05503-900 São Paulo, S.P., Brazil, and the §Department of Veterinary Science and Microbiology, University of Arizona, Tucson, Arizona 85721

Received for publication, December 29, 2003 , and in revised form, January 16, 2004.


   ABSTRACT
TOP
 ABSTRACT
INTRODUCTION
 EXPERIMENTAL PROCEDURES
 RESULTS AND DISCUSSION
 REFERENCES
 
Bites by Loxosceles spiders can produce severe clinical symptoms, including dermonecrosis, thrombosis, vascular leakage, hemolysis, and persistent inflammation. The causative factor is a sphingomyelinase D (SMaseD) that cleaves sphingomyelin into choline and ceramide 1-phosphate. A similar enzyme, showing comparable bioactivity, is secreted by certain pathogenic corynebacteria and acts as a potent virulence factor. However, the molecular basis for SMaseD toxicity is not well understood, which hampers effective therapy. Here we show that the spider and bacterial SMases D hydrolyze albumin-bound lysophosphatidylcholine (LPC), but not sphingosylphosphorylcholine, with Km values (~20–40 µM) well below the normal LPC levels in blood. Thus, toxic SMases D have intrinsic lysophospholipase D activity toward LPC. LPC hydrolysis yields the lipid mediator lysophosphatidic acid (LPA), a known inducer of platelet aggregation, endothelial hyperpermeability, and pro-inflammatory responses. Introduction of LPA1 receptor cDNA into LPA receptor-negative cells renders non-susceptible cells susceptible to SmaseD, but only in LPC-containing media. Degradation of circulating LPC to LPA with consequent activation of LPA receptors may have a previously unappreciated role in the pathophysiology of secreted SMases D.


   INTRODUCTION
TOP
 ABSTRACT
 INTRODUCTION
EXPERIMENTAL PROCEDURES
 RESULTS AND DISCUSSION
 REFERENCES
 
Envenomation by Loxosceles spiders, endemic to temperate and (sub)tropical regions of the Americas, Africa, and Europe, can lead to local skin injury as well as to serious systemic toxicity, including thrombus formation, vascular leakage, hemolysis, and persistent inflammation (1–3). In severe cases, the hematologic complications can lead to renal failure and death, especially in children (2, 3). Treatment is difficult; antivenoms are not very effective, and the use of corticosteroids or anti-inflammatory medication is controversial (3). The toxin responsible for the local and systemic effects of Loxosceles venom is an unusual sphingomyelinase D (SMaseD)1 that converts sphingomyelin (SM) in the outer leaflet of the plasma membrane to ceramide 1-phosphate (N-acylsphingosine 1-phosphate) (4–7). Strikingly, while SMaseD is not found elsewhere in the animal kingdom, a similar enzyme is produced as an exotoxin by some pathogenic bacteria, notably Corynebacterium pseudotuberculosis, Corynebacterium ulcerans, and Arcanobacterium (formerly Corynebacterium) hemolyticum (8–10). C. pseudotuberculosis causes lymphadenitis in animals but is also pathogenic for humans, while C. ulcerans and A. hemolyticum are pathogens of pharyngitis and other human infections (11); in no case is the molecular basis for virulence known (12). The SMaseD from C. pseudotuberculosis, also named SM-specific phospholipase D (PLD), is an essential virulence determinant that contributes to the persistence and spread of the bacteria within the host (13). The Loxosceles and C. pseudotuberculosis SMases D have the same molecular mass (31–32 kDa) and share about 30% sequence similarity (see "Results"). In model systems, the spider and bacterial enzymes provoke remarkably similar pathophysiological effects, including platelet aggregation, endothelial hyperpermeability, complement-dependent hemolysis, and neutrophil-dependent skin necrosis (4–7, 9, 14–16).

Despite decades of study it remains unclear how SMaseD can elicit such a wide variety of biological effects, particularly, since ceramide 1-phosphate is not known as a signaling molecule. In contrast to ceramide, which may reorganize lipid microdomains and associated signaling complexes (17, 18), ceramide 1-phosphate is a bilayer-preferring phospholipid that is unlikely to significantly perturb membrane structure. Furthermore, mammalian cells treated with SMaseD from either Loxosceles deserta or C. pseudotuberculosis do not convert newly formed ceramide 1-phosphate to ceramide nor does SMaseD treatment affect membrane permeability or cell viability (19, 20).

Given the lack of understanding of SMaseD bioactivity, we set out to re-examine the substrate specificity and cellular effects of the enzyme. Our interest was stirred by a report of more than 30 years ago, showing that partially purified SMaseD from C. pseudotuberculosis (ovis) can catalyze the release of choline from lysophosphatidylcholine (LPC) but not from phosphatidylcholine (PC) (21). LPC is an abundant plasma component and removal of its choline headgroup yields lysophosphatidic acid (LPA), now known as a pleiotropic lipid mediator acting on specific G protein-coupled receptors in numerous cell types (22, 23). Yet, the possibility that degradation of plasma LPC might contribute to SMaseD toxicity has received little attention to date. On the other hand, the reported Km value for LPC was very high, ~8 mM (21), suggesting that LPC is not a physiological substrate; moreover, LPC hydrolysis by partially purified SMaseD preparations could well be due to contaminating lyso-PLD activity.

In the present study, we demonstrate that the spider and bacterial SMases D have intrinsic lyso-PLD activity toward albumin-bound LPC and that expression of functional LPA receptors is necessary and sufficient for at least some of the biological responses to SMaseD. Our results suggest that degradation of circulating LPC to LPA, with consequent activation of LPA receptors in cells of the circulatory and vascular systems, may significantly contribute to SMaseD-mediated pathogenicity.


   EXPERIMENTAL PROCEDURES
TOP
 ABSTRACT
 INTRODUCTION
 EXPERIMENTAL PROCEDURES
RESULTS AND DISCUSSION
 REFERENCES
 
Cell Culture and Materials—Cells were grown in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal calf serum. Cells were exposed to serum-free DMEM for 24 h prior to experimentation, unless indicated otherwise. All phospholipids were obtained from Avanti Polar Lipids Inc. (Alabaster, AL) at the highest purity grade available. Fatty acid-free bovine serum albumin and monoclonal anti-phospho-MAP kinase antibody were from Sigma. Secondary antibodies (rabbit anti-mouse and swine anti-rabbit) conjugated to horseradish peroxidase were from Dako (Glostrup, Denmark). Highly purified PLD (57 kDa) from S. chromofuscus (24) was kindly provided by Dr. Mary F. Roberts (Boston College, Chestnut Hill, MA).

Recombinant SMaseD from Loxosceles laeta—Recombinant SMaseD (SMase I) from L. laeta was produced as described previously (7). In brief, mature L. laeta enzyme was expressed in Escherichia coli as a fusion protein, including a His6-tag at the N terminus and a 4-amino acid linker. The cells were collected by centrifugation and the bacterial pellet was resuspended in extraction buffer (300 mM NaCl, 100 mM Tris-HCl, pH 8.0) and disrupted by French pressure. The supernatant was loaded onto a Ni2+-chelating Sepharose Fast Flow column (Amersham Biosciences, Sweden, 1.0 x 6.4 cm), and the recombinant protein was eluted with buffer (300 mM NaCl, 100 mM Tris-HCl, pH 8.0, 0.8 M imidazole). Fractions of 1 ml were collected and analyzed by SDS-PAGE. Stock solutions were prepared in PBS at 1.0 mg of protein/ml.

Recombinant SMaseD from C. pseudotuberculosis—Recombinant C. pseudotuberculosis SMaseD was expressed in E. coli as a fusion protein composed of the mature enzyme with a 33-amino acid N-terminal extension containing a His6-tag (10). Recombinant enzyme was purified from the soluble fraction of cell lysates on TALON metal affinity resin (Clontech). SMaseD was eluted from the resin in 20 mM Tris-HCl, 100 mM NaCl, 100 mM imidazole at greater than 95% purity.

Choline Release Assay—SMaseD/PLD enzymatic activity was estimated by determining choline liberated from exogenously added phospholipid substrates, using a fluorimetric assay modified from Tokumura et al. (25). In the standard assay, the substrate was diluted in 100 µl of DMEM or HEPES-buffered saline (HBS; 140 mM NaCl, 5 mM KCl, 1 mM CaCl2, 1 mM MgCl2, 10 mM Hepes, pH 7.4). SM and PC substrates were applied as liposomes, whereas the lysophospholipids were complexed to fatty acid-free bovine serum albumin (5 mg/ml). After SMaseD/PLD addition, the reaction was left to proceed for 20 min at 37 °C. By adding 10 µl of a second assay mixture, the liberated free choline was oxidized (in 10 min) to betaine, and the H2O2 concomitantly generated was determined by fluorimetry. The second reaction mixture consisted of 1 unit/ml choline oxidase (Sigma), 0.06 unit/ml horseradish peroxidase, and 50 µM 3-(4-hydroxy-phenyl)propionic acid in HBS. The second reaction was left to proceed for 10 min. Fluorescence of the oxidized substrate was measured at an excitation of 320 nm and emission of 405 nm using a 96-well plate reader. SMaseD-induced choline release from LPC proceeded at a constant rate for at least 1 h at 37 °C, with the rate being proportional to the enzyme concentration.

LPA1 cDNA and Retroviral Transduction—Human LPA1 cDNA (GenBankTM accession number U78192 [GenBank] ) was amplified by PCR using primers carrying 5' XhoI + HA/3' NotI sites. N-terminally HA-tagged and C-terminally green fluorescent protein (GFP)-fused LPA1 receptor cDNAs were cloned into retroviral LZRS-IRES-Neo. Recombinant retrovirus produced in Phoenix packaging cells was used to infect rat B103 neuroblastoma cells or human HEK293 cells, essentially as described previously (26). After 48 h, transduced cells were selected in medium containing 1.0 mg/ml G418. Correct expression of LPA1 was confirmed by Western blotting and immunofluorescence.

LPA Receptor Internalization—HEK293 cells stably transfected with GFP-LPA1 receptor cDNA were fixed in 3.7% formaldehyde in PBS. After treatment with agonist, cells were washed and coverslips were mounted with vectashield (Vector Laboratories Inc., Burlingame, CA). GFP-LPA1 receptors were visualized by confocal microscopy.


   RESULTS AND DISCUSSION
TOP
 ABSTRACT
 INTRODUCTION
 EXPERIMENTAL PROCEDURES
 RESULTS AND DISCUSSION
REFERENCES
 
The secreted SMases D from L. laeta (Lox-SMaseD) (7) and C. pseudotuberculosis (Cp-SMaseD) (10) have the same molecular mass (31–32 kDa) and share 32% sequence similarity and 20% identity, including a conserved N-terminal histidine residue required for the enzymatic activity of Cp-SMaseD (see Supplemental Fig. 1, for sequence alignment, and Ref. 27). Despite being de facto PLDs, both SMases D are unique in that they show no homology to other phospholipid-metabolizing enzymes, and along with the broad specificity PLD from S. chromofuscus (28), they lack the conserved HKD sequence motif that characterizes the PLD superfamily (29).

We confirmed that the recombinant Lox- and Cp-SMases D catalyze the release of choline from SM but not PC, whereas the unrelated PLD from S. chromofuscus (Sc-PLD) cleaves both SM and PC (Fig. 1A). The intrinsic lyso-PLD activity of the recombinant SMases D was then assessed using 1-oleoyl-LPC and sphingosylphosphorylcholine (SPC or lyso-SM) as substrates; SPC is a normal plasma constituent (30) that would yield the potent lipid mediator sphingosine 1-phosphate following choline release. As shown in Fig. 1 (B and C), the spider and bacterial SMases D were both capable of liberating choline from albumin-bound LPC in physiological medium, thereby producing LPA (as confirmed by thin layer chromatography analysis). Surprisingly, SPC did not serve as a substrate for either SMaseD, even when SPC was applied at supra-physiological concentrations (50 µM, i.e. 3 orders of magnitude above normal plasma levels (30)) (Fig. 1B; only at still higher concentrations, some hydrolysis of SPC by Cp-SMaseD was observed, ~10% of that for LPC; data not shown). We further observed that both SMases D were capable of hydrolyzing naturally occurring ether-linked LPC (1-O-hexadecylglycero-3-phosphocholine or "lyso-platelet-activating factor") thereby producing alkyl-LPA, the most potent platelet-activating form of LPA (31); the unnatural alkyl-lysophospholipid Et-18-OCH(3) (1-octadecyl-2-methylglycero-3-phosphocholine; 50 µM) was not hydrolyzed by either SMaseD nor did it act as an inhibitor (data not shown). Many secreted phospholipases, including Sc-PLD and mammalian lyso-PLD (25), are dependent on calcium for their activity. In contrast, SMaseD-induced LPC hydrolysis has an absolute requirement for magnesium rather than calcium, since enzyme activity was abolished in the presence of EDTA but not EGTA, when assayed in buffer containing Ca2+ and Mg2+ as the only divalent metal ions (Fig. 1C).



View larger version (19K):
[in this window]
[in a new window]
  		FIG. 1.
Choline release from phospholipids induced by recombinant SMaseD from L. laeta and C. pseudotuberculosis. Choline release was determined fluorimetrically in HEPES-buffered DMEM, pH 7.4, at 37 °C (see "Experimental Procedures"). Lox-SMaseD and Cp-SMaseD were used at 10 nM. The nonspecific PLD from S. chromofuscus (Sc-PLD; 10 nM) was used as a positive control. Qualitatively similar results were obtained with enzyme concentrations up to 100 nM. Control denotes no enzyme addition. a, hydrolysis of PC and SM, added at 50 µM as multilamellar vesicles (liposomes). b, hydrolysis of LPC (1-oleoyl) and SPC, added at 50 µM and complexed to fatty acid-free albumin (5 mg/ml). c, dependence of LPC hydrolysis on the presence of EDTA or EGTA (4 mM). Error bars represent S.E. of the mean (n = 5).

 
By varying the LPC concentration, we determined that the spider and bacterial SMases D have an apparent Km value for 1-oleoyl-LPC of 44.4 ± 2.5 µM and 21.8 ± 3.0 µM, respectively (Fig. 2). Of note, these Km values are severalfold lower than the physiological LPC levels in human plasma (approximately 150 µM; (32, 33)), implying that LPC serves as a bona fide physiological substrate. For Lox-SMaseD, the apparent Vmax for LPC was 212 ± 6 nmol/min/mg of protein; for Cp-SMaseD, the Vmax value was 68 ± 2 nmol/min/mg of protein (Fig. 2). Taken together, these results indicate that the spider and bacterial SMases D possess Mg2+-dependent lyso-PLD activity toward acyl- and alkyl-LPC, but not toward SPC, with both enzymes showing similar catalytic efficiency (Vmax/Km, if Vmax is considered an approximation of kcat). For comparison, the estimated efficiency of SMaseD for LPC hydrolysis is approximately 1 order of magnitude lower than that of mammalian lyso-PLD or "autotaxin" (Km = 250 µM and Vmax = 9 mmol/min/mg (34)).



View larger version (15K):
[in this window]
[in a new window]
  		FIG. 2.
Saturation kinetics of SMaseD from L. laeta (a) and C. pseudotuberculosis (b). Rates of choline release from 1-oleoyl-LPC are plotted against increasing concentration of 1-oleoyl-LPC. Assays were carried out in HEPES-buffered DMEM (pH 7.4) at 37 °C (see "Experimental Procedures"). Data were fitted to the Michaelis-Menten equation, yielding the indicated apparent Km values. The Vmax values were 212 ± 6 nmol/min/mg for Lox-SMaseD and 66 ± 2 nmol/min/mg for Cp-SMaseD. Data points are the mean of three independent experiments each performed in triplicate. Error bars represent S.E. values.

 
We next examined the responsiveness of mammalian cells to SMaseD. While the nonspecific Sc-PLD can trigger diverse signaling events (35, 36), the SMases D failed to induce cellular responses under the same conditions. When preincubated with albumin-LPC complexes, however, the SMases D exhibited biological activity as evidenced by the induction of MAP kinase activity and cellular shape changes, consistent with SMaseD hydrolyzing extracellular LPC to receptor-active LPA. To prove this directly, we used rat B103 neuroblastoma cells that lack endogenous LPA receptors. Fig. 3 shows that SMaseD failed to activate MAP kinase (ERK1/2) in native B103 cells, either in the presence or absence of albumin-LPC. After forced expression of the prototypic LPA1 receptor in these cells, both SMases D mimicked LPA in activating MAP kinase, but only when preincubated with albumin-LPC (Fig. 3). Likewise, SMaseD mimicked LPA in inducing rapid cytoskeletal contraction in N1E-115 neuroblastoma cells, a typical RhoA-mediated response that also underlies LPA-induced endothelial barrier dysfunction (22, 37); but again, SMaseD acted only in the presence of albumin-LPC (see Supplemental Fig. 2).



View larger version (32K):
[in this window]
[in a new window]
  		FIG. 3.
LPA1 receptor expression confers cellular susceptibility to SMaseD. Detection of activated MAP kinase (ERK1/2) in LPA receptor-deficient B103 neuroblastoma cells and LPA1 receptor-expressing B103-LPA1 cells (26), as determined by using phospho-specific antibodies. Cells in serum-free DMEM were treated for 5 min with the indicated enzymes (20 nM) or 1-oleoyl-LPA (1 µM), in the presence or absence of 1-oleoyl-LPC (10 µM; complexed to 5 mg/ml fatty acid-free albumin).

 
Ligand activation of transmembrane receptors causes their internalization from the cell surface. As illustrated in Fig. 4, LPA triggers rapid internalization of its cognate receptors in stably transfected HEK293 cells; at 20–30 min following LPA addition, nearly all receptors were found intracellularly. In contrast, SMaseD alone did not affect LPA receptor localization in LPC-free medium. Upon incubation in albumin/LPC-containing medium, however, SMaseD induced efficient LPA receptor internalization albeit with somewhat slower kinetics than those observed with LPA (Fig. 4). Collectively, these results indicate that (i) SMaseD-mediated hydrolysis of albumin-bound LPC yields receptor-active LPA; and (ii) LPA receptor expression is necessary and sufficient for cellular susceptibility to SMaseD, provided that the cellular microenvironment contains albumin-LPC.



View larger version (59K):
[in this window]
[in a new window]
  		FIG. 4.
LPA1 receptor internalization induced by SMaseD. Subcellular localization of LPA1-GFP receptors stably expressed in HEK293 cells. GFP fluorescence is represented in gray-black for better contrast. Note complete LPA1 receptor internalization induced by 1-oleoyl-LPA (1 µM) after 10 min. Bacterial SMaseD/PLD induces receptor internalization only when preincubated with albumin-LPC (10 µM). Similar results were obtained with Lox-SMaseD.

 
Thus, when SMaseD comes into contact with blood, it will provoke the hydrolysis of albumin-bound LPC to generate LPA. Through activation of its cognate G protein-coupled receptors, LPA evokes a host of biological and pathophysiological responses in numerous cell types, including those of the circulatory and vascular systems (22, 23, 38). LPA levels in whole blood are normally very low, but the lipid accumulates during blood clotting to promote wound healing processes (39, 40). However, aberrant accumulation of LPA in blood may have deleterious effects, including endothelial barrier dysfunction (37), intravascular coagulation, and infiltration of inflammatory cells such as neutrophils (31, 38, 41, 42), responses that are strongly reminiscent of those to SMaseD in vivo. It is also of note that LPA stimulates Ca2+ entry into human erythrocytes (43) with subsequent exposure of phosphatidylserine (PS) on the cell surface.2 SMaseD likewise induces PS exposure in erythrocytes and this apparent loss of plasma membrane asymmetry is thought to contribute to complement-dependent hemolysis (16). The potential role of lyso-PLD activity in the hemolytic action of SMaseD obviously warrants further study.

Unlike envenomation by a Loxosceles spider bite, infections such as lymphadenitis caused by C. pseudotuberculosis result not only from the toxic effects of SMaseD per se but also from bacterial dissemination to host tissues such as the lymph nodes. A SMaseD-deficient mutant of C. pseudotuberculosis is unable to disseminate from the site of inoculation (13), and a popular theory is that vascular hyperpermeability caused by SMaseD, shown here to be likely a result of LPA production, aids in the escape of bacteria into the lymphatic system and subsequent spread to regional lymph nodes (44).

In conclusion, by building on early but largely overlooked evidence (21), we have demonstrated here that spider and bacterial SMases D have intrinsic lyso-PLD activity toward albumin-bound LPC and thereby generate bioactive LPA. To be consistent with phospholipase terminology, SMaseD should therefore be renamed "SM- and LPC-specific PLD." Our data thus provide, at least in part, a mechanistic explanation for the multiple biological responses to SMaseD observed in vivo. To what extent the hydrolysis of SM to ceramide 1-phosphate in the plasma membrane of target cells may cooperate with the concurrent generation of extracellular LPA to mediate pathogenicity remains an open question, although a recent report suggests that ceramide 1-phosphate may indirectly activate cytosolic phospholipase A2 leading to arachidonic acid release (45). Whatever the precise role of ceramide 1-phosphate, the finding that LPA receptor deficiency protects (nucleated) cells against SMaseD suggests that LPA receptors are potential targets in the treatment of Loxosceles envenomation as well as certain corynebacterial infections.


   FOOTNOTES
 
* This work was supported by the Dutch Cancer Society. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. Back

The on-line version of this article (available at http://www.jbc.org) contains Supplemental Figs. 1 and 2. Back

To whom correspondence should be addressed: Division of Cellular Biochemistry, The Netherlands Cancer Inst., Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands. Tel.: 31-20-5121971; Fax: 31-20-5121989; E-mail: w.moolenaar{at}nki.nl.

1 The abbreviations used are: SMaseD, sphingomyelinase D; LPC, lysophosphatidylcholine; LPA, lysophosphatidic acid; PC, phosphatidylcholine; SM, sphingomyelin; SPC, sphingosylphosphorylcholine; PLD, phospholipase D; MAP, mitogen-activated protein; GFP, green fluorescent protein; Lox-SMaseD, SMaseD from L. laeta; Cp-SMaseD, SMaseD from C. pseudotuberculosis; Sc-PLD, PLD from S. chromofuscus; DMEM, Dulbecco's modified Eagle's medium; HBS, HEPES-buffered saline; HA, hemagglutinin; PS, phosphatidylserine. Back

2 K. de Jong and F. A. Kuypers, personal communication. Back


   ACKNOWLEDGMENTS
 
We thank Paula Ruurs, Rafael Bernad, and Trudi Hengeveld for excellent technical assistance and Dr. Mary F. Roberts for providing Sc-PLD.



   REFERENCES
TOP
 ABSTRACT
 INTRODUCTION
 EXPERIMENTAL PROCEDURES
 RESULTS AND DISCUSSION
 REFERENCES
 

  1. Rees, R. S., O'Leary, J. P., and King, L. E., Jr. (1983) J. Surg. Res. 35, 1–10[CrossRef][Medline] [Order article via Infotrieve]
  2. Futrell, J. M. (1992) Am. J. Med. Sci. 304, 261–267[Medline] [Order article via Infotrieve]
  3. Sams, H. H., Dunnick, C. A., Smith, M. L., and King, L. E., Jr. (2001) J. Am. Acad. Dermatol. 44, 561–573[CrossRef][Medline] [Order article via Infotrieve]
  4. Forrester, L. J., Barrett, J. T., and Campbell, B. J. (1978) Arch. Biochem. Biophys. 187, 355–365[CrossRef][Medline] [Order article via Infotrieve]
  5. Kurpiewski, G., Forrester, L. J., Barrett, J. T., and Campbell, B. J. (1981) Biochim. Biophys. Acta 678, 467–476[Medline] [Order article via Infotrieve]
  6. Tambourgi, D. V., Magnoli, F. C., Van Den Berg, C. W., Morgan, B. P., de Araujo, P. S., Alves, E. W., and Da Silva, W. D. (1998) Biochem. Biophys. Res. Commun. 251, 366–373[CrossRef][Medline] [Order article via Infotrieve]
  7. Fernandes Pedrosa Mde, F., Junqueira de Azevedo Ide, L., Goncalves-de-Andrade, R. M., Van Den Berg, C. W., Ramos, C. R., Ho, P. L., and Tambourgi, D. V. (2002) Biochem. Biophys. Res. Commun. 298, 638–645[CrossRef][Medline] [Order article via Infotrieve]
  8. Soucek, A., Michalec, C., and Souckova, A. (1967) Biochim. Biophys. Acta 144, 180–182[Medline] [Order article via Infotrieve]
  9. Truett, A. P., III, and King, L. E., Jr. (1993) Adv. Lipid Res. 26, 275–291[Medline] [Order article via Infotrieve]
  10. McNamara, P. J., Cuevas, W. A., and Songer, J. G. (1995) Gene (Amst.) 156, 113–118[CrossRef][Medline] [Order article via Infotrieve]
  11. Lipsky, B. A., Goldberger, A. C., Tompkins, L. S., and Plorde, J. J. (1982) Rev. Infect. Dis. 4, 1220–1235[Medline] [Order article via Infotrieve]
  12. Songer, J. G. (1997) Trends Microbiol. 5, 156–161[CrossRef][Medline] [Order article via Infotrieve]
  13. McNamara, P. J., Bradley, G. A., and Songer, J. G. (1994) Mol. Microbiol. 12, 921–930[Medline] [Order article via Infotrieve]
  14. Carne, H. R., and Onon, E. O. (1978) Nature 271, 246–248[CrossRef][Medline] [Order article via Infotrieve]
  15. Bernheimer, A. W., Campbell, B. J., and Forrester, L. J. (1985) Science 228, 590–591[Abstract/Free Full Text]
  16. Tambourgi, D. V., Sousa Da Silva, M., Billington, S. J., Goncalves De Andrade, R. M., Magnoli, F. C., Songer, J. G., and Van Den Berg, C. W. (2002) Immunology 107, 93–101[CrossRef][Medline] [Order article via Infotrieve]
  17. Gulbins, E., and Kolesnick, R. (2003) Oncogene 22, 7070–7077[CrossRef][Medline] [Order article via Infotrieve]
  18. van Blitterswijk, W. J., Van Der Luit, A. H., Veldman, R. J., Verheij, M., and Borst, J. (2003) Biochem. J. 369, 199–211[CrossRef][Medline] [Order article via Infotrieve]
  19. Feldhaus, M. J., Weyrich, A. S., Zimmerman, G. A., and McIntyre, T. M. (2002) J. Biol. Chem. 277, 4285–4293[Abstract/Free Full Text]
  20. Subbaiah, P. V., Billington, S. J., Jost, B. H., Songer, J. G., and Lange, Y. (2003) J. Lipid Res. 44, 1574–1580[Abstract/Free Full Text]
  21. Soucek, A., Michalec, C., and Souckova, A. (1971) Biochim. Biophys. Acta 227, 116–128[Medline] [Order article via Infotrieve]
  22. Moolenaar, W. H. (1999) Exp. Cell Res. 253, 230–238[CrossRef][Medline] [Order article via Infotrieve]
  23. Chun, J., Goetzl, E. J., Hla, T., Igarashi, Y., Lynch, K. R., Moolenaar, W., Pyne, S., and Tigyi, G. (2002) Pharmacol. Rev. 54, 265–269[Abstract/Free Full Text]
  24. Geng, D., Chura, J., and Roberts, M. F. (1998) J. Biol. Chem. 273, 12195–12202[Abstract/Free Full Text]
  25. Tokumura, A., Majima, E., Kariya, Y., Tominaga, K., Kogure, K., Yasuda, K., and Fukuzawa, K. (2002) J. Biol. Chem. 277, 39436–39442[Abstract/Free Full Text]
  26. Van Leeuwen, F. N., Olivo, C., Grivell, S., Giepmans, B. N., Collard, J. G., and Moolenaar, W. H. (2003) J. Biol. Chem. 278, 400–406[Abstract/Free Full Text]
  27. Tachedjian, M., Krywult, J., Moore, R. J., and Hodgson, A. L. (1995) Vaccine 13, 1785–1792[CrossRef][Medline] [Order article via Infotrieve]
  28. Yang, H., and Roberts, M. F. (2002) Protein Sci. 11, 2958–2968[CrossRef][Medline] [Order article via Infotrieve]
  29. Exton, J. H. (2002) FEBS Lett. 531, 58–61[CrossRef][Medline] [Order article via Infotrieve]
  30. Liliom, K., Sun, G., Bunemann, M., Virag, T., Nusser, N., Baker, D. L., Wang, D. A., Fabian, M. J., Brandts, B., Bender, K., Eickel, A., Malik, K. U., Miller, D. D., Desiderio, D. M., Tigyi, G., and Pott, L. (2004) Biochem. J. 355, 189–197
  31. Haserueck, N., Erl, W., Pandey, D., Tigyi, G., Ohlmann, P., Ravanat, C., Gachet, C., and Siess, W. (2004) Blood, in press
  32. Subbaiah, P. V., Chen, C. H., Bagdade, J. D., and Albers, J. J. (1985) J. Biol. Chem. 260, 5308–5314[Abstract/Free Full Text]
  33. Croset, M., Brossard, N., Polette, A., and Lagarde, M. (2000) Biochem. J. 345, 61–67
  34. Umezu-Goto, M., Kishi, Y., Taira, A., Hama, K., Dohmae, N., Takio, K., Yamori, T., Mills, G. B., Inoue, K., Aoki, J., and Arai, H. (2002) J. Cell Biol. 158, 227–233[Abstract/Free Full Text]
  35. Moolenaar, W. H., Kruijer, W., Tilly, B. C., Verlaan, I., Bierman, A. J., and de Laat, S. W. (1986) Nature 323, 171–173[CrossRef][Medline] [Order article via Infotrieve]
  36. van Dijk, M. C., Postma, F., Hilkmann, H., Jalink, K., van Blitterswijk, W. J., and Moolenaar, W. H. (1998) Curr. Biol. 8, 386–392[CrossRef][Medline] [Order article via Infotrieve]
  37. Nieuw Amerongen, G. P., Vermeer, M. A., and Van Hinsbergh, V. W. (2000) Arterioscler. Thromb. Vasc. Biol. 20, E127–E133
  38. Siess, W. (2002) Biochim. Biophys. Acta 1582, 204–215[Medline] [Order article via Infotrieve]
  39. Eichholtz, T., Jalink, K., Fahrenfort, I., and Moolenaar, W. H. (1993) Biochem. J. 291, 677–680
  40. Sano, T., Baker, D., Virag, T., Wada, A., Yatomi, Y., Kobayashi, T., Igarashi, Y., and Tigyi, G. (2002) J. Biol. Chem. 277, 21197–21206[Abstract/Free Full Text]
  41. Fueller, M., Wang de, A., Tigyi, G., and Siess, W. (2003) Cell Signal. 15, 367–375[CrossRef][Medline] [Order article via Infotrieve]
  42. Hashimoto, T., Yamashita, M., Ohata, H., and Momose, K. (2003) J. Pharmacol. Sci. 91, 8–14[CrossRef][Medline] [Order article via Infotrieve]
  43. Yang, L., Andrews, D. A., and Low, P. S. (2000) Blood 95, 2420–2425[Abstract/Free Full Text]
  44. Batey, R. G. (1986) Aust. Vet. J. 63, 269–272[Medline] [Order article via Infotrieve]
  45. Pettus, B. J., Bielawska, A., Spiegel, S., Roddy, P., Hannun, Y. A., and Chalfant, C. E. (2003) J. Biol. Chem. 278, 38206–38213[Abstract/Free Full Text]

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
 Mol Cancer ResHome page
R. M. Kortlever, T. R. Brummelkamp, L. A. van Meeteren, W. H. Moolenaar, and R. Bernards
Suppression of the p53-Dependent Replicative Senescence Response by Lysophosphatidic Acid Signaling
Mol. Cancer Res., September 1, 2008; 6(9): 1452 - 1460.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
M. Murph, T. Tanaka, S. Liu, and G. B. Mills
Of Spiders and Crabs: The Emergence of Lysophospholipids and Their Metabolic Pathways as Targets for Therapy in Cancer.
Clin. Cancer Res., November 15, 2006; 12(22): 6598 - 6602.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
L. A. van Meeteren, P. Ruurs, C. Stortelers, P. Bouwman, M. A. van Rooijen, J. P. Pradere, T. R. Pettit, M. J. O. Wakelam, J. S. Saulnier-Blache, C. L. Mummery, et al.
Autotaxin, a Secreted Lysophospholipase D, Is Essential for Blood Vessel Formation during Development.
Mol. Cell. Biol., July 1, 2006; 26(13): 5015 - 5022.
[Abstract] [Full Text] [PDF]

Home page
 BioinformaticsHome page
M. H. J. Cordes and G. J. Binford
Lateral gene transfer of a dermonecrotic toxin between spiders and bacteria
Bioinformatics, February 1, 2006; 22(3): 264 - 268.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
D. A. Lin and J. A. Boyce
IL-4 Regulates MEK Expression Required for Lysophosphatidic Acid-Mediated Chemokine Generation by Human Mast Cells
J. Immunol., October 15, 2005; 175(8): 5430 - 5438.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
L. A. van Meeteren, P. Ruurs, E. Christodoulou, J. W. Goding, H. Takakusa, K. Kikuchi, A. Perrakis, T. Nagano, and W. H. Moolenaar
Inhibition of Autotaxin by Lysophosphatidic Acid and Sphingosine 1-Phosphate
J. Biol. Chem., June 3, 2005; 280(22): 21155 - 21161.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. T. Murakami, M. F. Fernandes-Pedrosa, D. V. Tambourgi, and R. K. Arni
Structural Basis for Metal Ion Coordination and the Catalytic Mechanism of Sphingomyelinases D
J. Biol. Chem., April 8, 2005; 280(14): 13658 - 13664.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Supplemental Data
Right arrow All Versions of this Article:
279/12/10833    most recent
C300563200v1
Right arrow Submit a Letter to Editor
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
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 arrowRequest Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by van Meeteren, L. A.
Right arrow Articles by Moolenaar, W. H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by van Meeteren, L. A.
Right arrow Articles by Moolenaar, W. H.
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   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2004 by the American Society for Biochemistry and Molecular Biology.
Advertisement
spacer
Advertisement
Advertisement