The sphingolipid degradation product trans-2-hexadecenal forms adducts with DNA

doi:10.1016/j.bbrc.2012.06.012

Biochemical and Biophysical Research Communications

Volume 424, Issue 1, 20 July 2012, Pages 18-21

https://doi.org/10.1016/j.bbrc.2012.06.012 Get rights and content

Abstract

Sphingosine 1-phosphate, a bioactive signaling molecule with diverse cellular functions, is irreversibly degraded by the endoplasmic reticulum enzyme sphingosine 1-phosphate lyase, generating trans-2-hexadecenal and phosphoethanolamine. We recently demonstrated that trans-2-hexadecenal causes cytoskeletal reorganization, detachment, and apoptosis in multiple cell types via a JNK-dependent pathway. These findings and the known chemistry of related α,β-unsaturated aldehydes raise the possibility that trans-2-hexadecenal may interact with additional cellular components. In this study, we show that it reacts readily with deoxyguanosine and DNA to produce the diastereomeric cyclic 1,N²-deoxyguanosine adducts 3-(2-deoxy-β-d-erythro-pentofuranosyl)-5,6,7,8-tetrahydro-8R-hydroxy-6R-tridecylpyrimido[1,2-a]purine-10(3H)one and 3-(2-deoxy-β-d-erythro-pentofuranosyl)-5,6,7,8-tetrahydro-8S-hydroxy-6S-tridecylpyrimido[1,2-a]purine-10(3H)one. Thus, our findings suggest that trans-2-hexadecenal produced endogenously by sphingosine 1-phosphate lyase can react directly with DNA forming aldehyde-derived DNA adducts with potentially mutagenic consequences.

Highlights

► trans-2-Hexadecenal reacts with DNA to produce adducts. ► Structurally unusual lipophilic DNA adducts are formed. ► The adducts were characterized by NMR, MS, and UV.

Introduction

Sphingosine 1-phosphate lyase (SPL) catalyzes the conversion of sphingosine 1-phosphate (1) to trans-2-hexadecenal (2) and phosphoethanolamine (3) (Scheme 1) [1]. The roles of 1 in cell and animal physiology have been well described [2], but the biological consequences and fate of metabolic products of 1 such as 2 are not well understood. We have previously shown that 2 causes cytoskeletal reorganization, detachment, and apoptosis in multiple cell types via a JNK-dependent pathway, indicating that it may interact with multiple cellular components including DNA [3]. Previous studies have shown that α,β-unsaturated aldehydes such as acrolein, crotonaldehyde (2-butenal), and 4-hydroxy-2-nonenal react readily with deoxyguanosine (dG, 4) and DNA to produce cyclic 1,N²-propanodeoxyguanosine adducts with a variety of mutagenic properties [4], [5], [6], [7], [8], [9], [10]. Therefore, we hypothesized that 2 would undergo a similar reaction (Scheme 1), potentially producing an adduct such as 5, which could be central to some of the physiologic effects of 2. We tested this hypothesis in the study reported here.

Section snippets

HPLC systems

System 1 consisted of a Waters Corp. model 600 system controller, two model 501 pumps, a model 440 UV detector (254 nm), and an Agilent Technologies 1100 series auto-sampler. A Luna C₁₈(2) reversed-phase column (250 × 4.6 mm, 5 μm; Phenomenex) was used for the separation. Elution was performed with a linear gradient from 85% 15 mM NH₄OAc in CH₃OH to 100% CH₃OH over 30 min and held for 20 min. The flow rate was 0.7 ml/min.

System 2 used an Agilent Technologies 1100 capillary HPLC system equipped with a 5

Results and discussion

trans-2-Hexadecenal (2) was allowed to react with dG (4), and the products were analyzed by LC–ESI-MS/MS-SRM. The transitions monitored were m/z 506 → 390 [BH]⁺, m/z 390 → 346 [BH–CH₂CHOH]⁺, m/z 390 → 372 [BH–H₂O]⁺, m/z 390 → 190 [BH–(CH₃(CH₂)₁₂ + OH)]⁺, and m/z 390 → 152 [GH]⁺ (Scheme 2). As shown in Fig. 1, all of these transitions were observed and are consistent with the expected product, 5.

The UV spectrum and the 850 MHz ¹H NMR and ¹³C NMR spectra of 5 are completely consistent with those of other

Acknowledgments

This study was supported by NIH grants CA-77528 and CA-129438 (JDS), CA-81301 (SSH), and HL-083187 (RB). Mass spectrometry was carried out in the Analytical Biochemistry Shared Resource of the Masonic Cancer Center, supported in part by NIH grant CA-77598. We thank Todd Rappe, University of Minnesota NMR Center, for acquiring the NMR spectra.

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Biochemical and Biophysical Research Communications

Abstract

Highlights

Introduction

Section snippets

HPLC systems

Results and discussion

Acknowledgments

References (19)

The sphingolipid degradation product trans-2-hexadecenal induces cytoskeletal reorganization and apoptosis in a JNK-dependent manner

Cell. Signal

Oxidative cleavage of lipids with sodium metaperiodate in pyridine

J. Lipid Res.

Detection of the acrolein-derived cyclic DNA adduct by a quantitative ³²P-postlabeling/solid-phase extraction/HPLC method: blocking its artifact formation with glutathione

Anal. Biochem.

Sphingosine-1-phosphate metabolism and intestinal tumorigenesis: lipid signaling strikes again

Cell Cycle

Lyase to live by: sphingosine phosphate lyase as a therapeutic target

Expert Opin. Ther. Targets

Formation of cyclic 1,N²-propanodeoxyguanosine adducts in DNA upon reaction with acrolein or crotonaldehyde

Cancer Res.

A study of reactions of α,β-unsaturated carbonyl compounds with deoxyguanosine

J. Org. Chem.

Detection of 1,N²-propanodeoxyguanosine adducts as potential endogenous DNA lesions in rodent and human tissues

Cancer Res.

Role of 1,N²-propanodeoxyguanosine adducts as endogenous DNA lesions in rodents and humans

Cited by (25)

S1P and plasmalogen derived fatty aldehydes in cellular signaling and functions

A Mechanism-Based Sphingosine-1-phosphate Lyase Inhibitor

Sphingosine phosphate lyase insufficiency syndrome (SPLIS): A novel inborn error of sphingolipid metabolism

Sphingolipids and glycerophospholipids – The “ying and yang” of lipotoxicity in metabolic diseases

The sphingosine 1-phosphate breakdown product, (2E)-hexadecenal, forms protein adducts and glutathione conjugates in vitro

A facile stable-isotope dilution method for determination of sphingosine phosphate lyase activity

The sphingolipid degradation product trans-2-hexadecenal forms adducts with DNA

Abstract

Highlights

Introduction

Section snippets

HPLC systems

Results and discussion

Acknowledgments

Cell. Signal

J. Lipid Res.

Anal. Biochem.

Sphingosine-1-phosphate metabolism and intestinal tumorigenesis: lipid signaling strikes again

Cell Cycle

Lyase to live by: sphingosine phosphate lyase as a therapeutic target

Expert Opin. Ther. Targets

Formation of cyclic 1,N2-propanodeoxyguanosine adducts in DNA upon reaction with acrolein or crotonaldehyde

Cancer Res.

A study of reactions of α,β-unsaturated carbonyl compounds with deoxyguanosine

J. Org. Chem.

Detection of 1,N2-propanodeoxyguanosine adducts as potential endogenous DNA lesions in rodent and human tissues

Cancer Res.

Role of 1,N2-propanodeoxyguanosine adducts as endogenous DNA lesions in rodents and humans

Formation of cyclic 1,N²-propanodeoxyguanosine adducts in DNA upon reaction with acrolein or crotonaldehyde

Detection of 1,N²-propanodeoxyguanosine adducts as potential endogenous DNA lesions in rodent and human tissues

Role of 1,N²-propanodeoxyguanosine adducts as endogenous DNA lesions in rodents and humans