Identification and Characterization of a Novel Human Sphingosine-1-phosphate Phosphohydrolase, hSPP2

Ogawa, Chie; Kihara, Akio; Gokoh, Maiko; Igarashi, Yasuyuki

doi:10.1074/jbc.m209514200

Cited by 161 publications

(120 citation statements)

References 23 publications

(19 reference statements)

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“…Indirect immunofluorescence microscopy was performed as described previously [12]. For nile red staining, cells were incubated with 1 μg/ml nile red (Wako Pure Chemical Industries) for 30 min at room temperature, followed by blocking with BSA and staining with antibodies.…”

Section: Immunofluorescence Microscopymentioning

confidence: 99%

Identification of acyl-CoA synthetases involved in the mammalian sphingosine 1-phosphate metabolic pathway

Ohkuni

Ohno

Kihara

2013

Biochemical and Biophysical Research Communications

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2 Highlights• Eight ACSs exhibited activities toward the S1P metabolite trans-2-hecadecenoic acid.• ACS isozymes involved in S1P metabolism were localized in the ER.• Inhibition of ACSs caused accumulation of trans-2-hexadecenoic acid. 3 ABSTRACTSphingosine 1-phosphate (S1P) plays important roles both as a bioactive lipid molecule and an intermediate of the sphingolipid-to-glycerophospholipid metabolic pathway. To identify human acyl-CoA synthetases (ACSs) involved in S1P metabolism, we cloned all 26 human ACS genes and examined their abilities to restore deficient sphingolipid-to-glycerophospholipid metabolism in a yeast mutant lacking two ACS genes, FAA1 and FAA4. Here, in addition to the previously identified ACSL family members (ACSL1, 3, 4, 5, and 6), we found that ACSVL1, ACSVL4, and ACSBG1 also restored metabolism. All 8 ACSs were localized either exclusively or partly to the endoplasmic reticulum (ER), where S1P metabolism takes place. We previously proposed the entire S1P metabolic pathway from results obtained using yeast cells, i.e., S1P is metabolized totrans-2-hexadecenoyl-CoA, and palmitoyl-CoA. However, as S1P is not a naturally occurring long-chain base 1-phosphate in yeast, the validity of this pathway required further verification using mammalian cells. In the present study, we treated HeLa cells with the ACS inhibitor triacsin C and found that inhibition of ACSs resulted in accumulation of trans-2-hexadecenoic acid as in ACS mutant yeast. From these results, we conclude that S1P is metabolized by a common pathway in eukaryotes.

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Section: Immunofluorescence Microscopymentioning

confidence: 99%

Identification of acyl-CoA synthetases involved in the mammalian sphingosine 1-phosphate metabolic pathway

Ohkuni

Ohno

Kihara

2013

Biochemical and Biophysical Research Communications

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“…For example, secreted phosphoprotein 2 (Spp2) is a lipid modulatory gene involved in signaling of S1p and is reportedly important in the developing kidney (Kirby et al 2009). Although it is known to be expressed at high levels in the adult mouse kidney (BioGPS, GNF1M) and in the human heart and kidney (Ogawa et al 2003), its function and anatomical localization in the kidney have not been previously reported. We show here (Fig.…”

Section: Kidneymentioning

confidence: 99%

A lacZ reporter gene expression atlas for 313 adult KOMP mutant mouse lines

et al. 2015

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Expression of the bacterial beta-galactosidase reporter gene (lacZ) in the vector used for the Knockout Mouse Project (KOMP) is driven by the endogenous promoter of the target gene. In tissues from KOMP mice, histochemical staining for LacZ enzyme activity can be used to determine gene expression patterns. With this technique, we have produced a comprehensive resource of gene expression using both whole mount (WM) and frozen section (FS) LacZ staining in 313 unique KOMP mutant mouse lines. Of these, ∼80% of mutants showed specific staining in one or more tissues, while ∼20% showed no specific staining, ∼13% had staining in only one tissue, and ∼25% had staining in >6 tissues. The highest frequency of specific staining occurred in the brain (∼50%), male gonads (42%), and kidney (39%). The WM method was useful for rapidly identifying whole organ and some substructure staining, while the FS method often revealed substructure and cellular staining specificity. Both staining methods had >90% repeatability in biological replicates. Nonspecific LacZ staining occurs in some tissues due to the presence of bacteria or endogenous enzyme activity. However, this can be effectively distinguished from reporter gene activity by the combination of the WM and FS methods. After careful annotation, LacZ staining patterns in a high percentage of mutants revealed a unique structure-function not previously reported for many of these genes. The validation of methods for LacZ staining, annotation, and expression analysis reported here provides unique insights into the function of genes for which little is currently known.[Supplemental material is available for this article.]

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“…Sphingosine kinase (SphK) catalyzes the synthesis of S1P (2), which can be irreversibly degraded by S1P lyase (9 -11) and reversibly converted back to sphingosine by S1P phosphatases (SPPs) (12)(13)(14)(15). Recently, two isoforms, designated SPP-1 (12,13,16) and SPP-2 (15), have been identified.…”

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confidence: 99%

Role of Sphingosine-1-phosphate Phosphatase 1 in Epidermal Growth Factor-induced Chemotaxis

Stunff

Mikami

Giussani

et al. 2004

Journal of Biological Chemistry

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Sphingosine-1-phosphate (S1P) is the ligand for a family of specific G protein-coupled receptors that regulate a wide variety of cellular functions, including cytoskeletal rearrangements and cell motility. Because of the pivotal role of S1P, its levels are low and tightly regulated in a spatial-temporal manner through its synthesis catalyzed by sphingosine kinases and degradation by an S1P lyase and specific S1P phosphatases (SPP). Surprisingly, down-regulation of SPP-1 enhanced migration toward epidermal growth factor (EGF); conversely, overexpression of SPP-1, which is localized in the endoplasmic reticulum, attenuated migration toward EGF. To determine whether the inhibitory effect on EGF-induced migration was because of decreased S1P or increased ceramide as a consequence of acylation of increased sphingosine by ceramide synthase, we used fumonisin B1, a specific inhibitor of ceramide synthase. Although fumonisin B1 blocked ceramide production and increased sphingosine, it did not reverse the negative effect of SPP-1 expression on EGF-or S1P-induced chemotaxis. EGF activated the epidermal growth factor receptor to the same extent in SPP-1-expressing cells, yet ERK1/2 activation was impaired. In agreement, PD98059, an inhibitor of the ERK-activating enzyme MEK, decreased EGF-stimulated migration. We next examined the possibility that intracellularly generated S1P might be involved in activating a G protein-coupled S1P receptor important for EGF-directed migration. Treatment with pertussis toxin to inactivate G␣ i suppressed EGF-induced migration. Moreover, expression of regulator of G protein signaling 3, which inhibits S1P receptor signaling and completely prevented ERK1/2 activation mediated by S1P receptors, not only reduced migration toward S1P but also markedly reduced migration toward EGF. Collectively, these results suggest that metabolism of S1P by SPP-1 is important for EGF-directed cell migration.The bioactive sphingolipid, sphingosine-1-phosphate (S1P), 1 is a ligand for a family of five specific G protein-coupled receptors (S1P 1 , S1P 2 , S1P 3 , S1P 4 , and S1P 5 ) (1) that are coupled to various G proteins. These receptors regulate diverse cellular processes and have been implicated in cytoskeletal rearrangement and regulation of cell movement (2, 3). The founding member of this receptor family, S1P 1 , couples to a G i pathway (4), and its gene disruption in mice demonstrated an essential function for vascular maturation (5). More recently, several studies have established that S1P 1 is also essential for lymphocyte recirculation and that it regulates egress from peripheral lymphoid organs (6, 7). In addition to its extracellular functions, S1P acts intracellularly to regulate cell survival (8), yet its direct targets have not yet been elucidated.As with other potent lipid mediators, levels of S1P are low and tightly regulated in a spatial-temporal manner. Sphingosine kinase (SphK) catalyzes the synthesis of S1P (2), which can be irreversibly degraded by S1P lyase (9 -11) and reversibly conver...

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Identification and Characterization of a Novel Human Sphingosine-1-phosphate Phosphohydrolase, hSPP2

Cited by 161 publications

References 23 publications

Identification of acyl-CoA synthetases involved in the mammalian sphingosine 1-phosphate metabolic pathway

Identification of acyl-CoA synthetases involved in the mammalian sphingosine 1-phosphate metabolic pathway

A lacZ reporter gene expression atlas for 313 adult KOMP mutant mouse lines

Role of Sphingosine-1-phosphate Phosphatase 1 in Epidermal Growth Factor-induced Chemotaxis

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