Abstract
Interest in sphingolipids has increased in the past couple of decades as the number of biological activities identified has greatly expanded. These include roles in inflammation, proliferation, survival, and metastasis. Sphingolipids can exert these effects through an increasing number of identified interacting cellular targets. To facilitate the understanding of the intrinsic biology of sphingolipids and the development of sphingolipid-based therapeutics, further knowledge is needed. Various analytical protocols assist this endeavor, with mass spectrometry-based techniques seeing increasing usage, especially for measuring steady-state lipid levels. The area of mass spectrometry-based proteomics is also seeing increased usage in the study of lipid biology. This chapter provides an introduction to hypothesis-generating and hypothesis-testing protein-based analytical approaches to investigate sphingolipids and sphingolipid-metabolizing enzymes. These tools can serve to identify how sphingolipids regulate the proteome, to define how post-translational modifications control enzymatic activity, to identify protein–protein and protein–lipid interactions as well as to facilitate inhibitor development, among other concepts. These approaches can help delineate the roles and consequences of perturbations of sphingolipid metabolism in cancer.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Hartmann D et al (2012) Long chain ceramides and very long chain ceramides have opposite effects on human breast and colon cancer cell growth. Int J Biochem Cell Biol 44(4):620–628
Karahatay S et al (2007) Clinical relevance of ceramide metabolism in the pathogenesis of human head and neck squamous cell carcinoma (HNSCC): attenuation of C(18)-ceramide in HNSCC tumors correlates with lymphovascular invasion and nodal metastasis. Cancer Lett 256(1):101–111
Saddoughi SA et al (2011) Results of a phase II trial of gemcitabine plus doxorubicin in patients with recurrent head and neck cancers: serum C(1)(8)-ceramide as a novel biomarker for monitoring response. Clin Cancer Res 17(18):6097–6105
Alberg AJ et al (2013) Plasma sphingolipids and lung cancer: a population-based, nested case-control study. Cancer Epidemiol Biomarkers Prev 22(8):1374–1382
Jiang Y et al (2013) Altered sphingolipid metabolism in patients with metastatic pancreatic cancer. Biomolecules 3(3):435–448
Han X, Yang K, Gross RW (2012) Multi-dimensional mass spectrometry-based shotgun lipidomics and novel strategies for lipidomic analyses. Mass Spectrom Rev 31(1):134–178
Fillet M et al (2005) Differential expression of proteins in response to ceramide-mediated stress signal in colon cancer cells by 2-D gel electrophoresis and MALDI-TOF-MS. J Proteome Res 4(3):870–880
Renert AF et al (2009) The proapoptotic C16-ceramide-dependent pathway requires the death-promoting factor Btf in colon adenocarcinoma cells. J Proteome Res 8(10):4810–4822
Kota V et al (2013) 2’-hydroxy C16-ceramide induces apoptosis-associated proteomic changes in C6 glioma cells. J Proteome Res 12(10):4366–4367
Parent N et al (2009) Proteomic analysis of enriched lysosomes at early phase of camptothecin-induced apoptosis in human U-937 cells. J Proteomics 72(6):960–973
Kim SY et al (2009) Proteomic identification of proteins translocated to membrane microdomains upon treatment of fibroblasts with the glycosphingolipid, C8-beta-D-lactosylceramide. Proteomics 9(18):4321–4328
Everley RA et al (2013) Increasing throughput in targeted proteomics assays: 54-plex quantitation in a single mass spectrometry run. Anal Chem 85(11):5340–5346
McClatchy DB, Yates JR 3rd (2008) Stable isotope labeling in mammals (SILAM). Methods Mol Biol 1156:133–146
Pozniak Y, Geiger T (2014) Design and application of super-SILAC for proteome quantification. Methods Mol Biol 1188:281–291
Zhou F et al (2013) Genome-scale proteome quantification by DEEP SEQ mass spectrometry. Nat Commun 4:2171
Mann M et al (2002) Analysis of protein phosphorylation using mass spectrometry: deciphering the phosphoproteome. Trends Biotechnol 20(6):261–268
Lundby A et al (2012) Quantitative maps of protein phosphorylation sites across 14 different rat organs and tissues. Nat Commun 3:876
Hao P, Guo T, Sze SK (2011) Simultaneous analysis of proteome, phospho- and glycoproteome of rat kidney tissue with electrostatic repulsion hydrophilic interaction chromatography. PLoS One 6(2), e16884
Mertins P et al (2013) Integrated proteomic analysis of post-translational modifications by serial enrichment. Nat Methods 10(7):634–637
Gillet LC et al (2012) Targeted data extraction of the MS/MS spectra generated by data-independent acquisition: a new concept for consistent and accurate proteome analysis. Mol Cell Proteomics 11(6):O111.016717
Momin AA et al (2011) A method for visualization of “omic” datasets for sphingolipid metabolism to predict potentially interesting differences. J Lipid Res 52(6):1073–1083
Lundberg E et al (2010) Defining the transcriptome and proteome in three functionally different human cell lines. Mol Syst Biol 6:450
Schwanhausser B et al (2011) Global quantification of mammalian gene expression control. Nature 473(7347):337–342
Gerber SA et al (2003) Absolute quantification of proteins and phosphoproteins from cell lysates by tandem MS. Proc Natl Acad Sci U S A 100(12):6940–6945
Anderson NL et al (2004) Mass spectrometric quantitation of peptides and proteins using stable isotope standards and capture by anti-peptide antibodies (SISCAPA). J Proteome Res 3(2):235–244
Bereman MS et al (2012) The development of selected reaction monitoring methods for targeted proteomics via empirical refinement. Proteomics 12(8):1134–1141
Mohammed Y et al (2014) PeptidePicker: a scientific workflow with web interface for selecting appropriate peptides for targeted proteomics experiments. J Proteomics 106C:51–161
Wilhelm M et al (2014) Mass-spectrometry-based draft of the human proteome. Nature 509(7502):582–587
Kim MS et al (2014) A draft map of the human proteome. Nature 509(7502):575–581
Zeidan YH, Hannun YA (2007) Activation of acid sphingomyelinase by protein kinase Cdelta-mediated phosphorylation. J Biol Chem 282(15):11549–11561
Parent N et al (2011) Protein kinase C-delta isoform mediates lysosome labilization in DNA damage-induced apoptosis. Int J Oncol 38(2):313–324
Pitson SM et al (2003) Activation of sphingosine kinase 1 by ERK1/2-mediated phosphorylation. EMBO J 22(20):5491–5500
Franzen R et al (2002) Nitric oxide induces degradation of the neutral ceramidase in rat renal mesangial cells and is counterregulated by protein kinase C. J Biol Chem 277(48):46184–46190
Galadari S et al (2006) Identification of a novel amidase motif in neutral ceramidase. Biochem J 393(Pt 3):687–695
Tada E et al (2010) Activation of ceramidase and ceramide kinase by vanadate via a tyrosine kinase-mediated pathway. J Pharmacol Sci 114(4):420–432
Chen WQ et al (2010) Ceramide kinase profiling by mass spectrometry reveals a conserved phosphorylation pattern downstream of the catalytic site. J Proteome Res 9(1):420–429
Ferlinz K et al (2001) Human acid ceramidase: processing, glycosylation, and lysosomal targeting. J Biol Chem 276(38):35352–35360
Schulze H, Schepers U, Sandhoff K (2007) Overexpression and mass spectrometry analysis of mature human acid ceramidase. Biol Chem 388(12):1333–1343
Rodriguez J et al (2008) Does trypsin cut before proline? J Proteome Res 7(1):300–305
Stahelin RV et al (2005) The mechanism of membrane targeting of human sphingosine kinase 1. J Biol Chem 280(52):43030–43038
Pflieger D et al (2008) Quantitative proteomic analysis of protein complexes: concurrent identification of interactors and their state of phosphorylation. Mol Cell Proteomics 7(2):326–346
Glibert P et al (2015) Phospho-iTRAQ: assessing isobaric labels for the large-scale study of phosphopeptide stoichiometry. J Proteome Res 14(2):839–849
Frese CK et al (2011) Improved peptide identification by targeted fragmentation using CID, HCD and ETD on an LTQ-Orbitrap Velos. J Proteome Res 10(5):2377–2388
Kumagai K, Kawano-Kawada M, Hanada K (2014) Phosphoregulation of the ceramide transport protein CERT at serine 315 in the interaction with VAMP-associated protein (VAP) for inter-organelle trafficking of ceramide in mammalian cells. J Biol Chem 289(15):10748–10760
Cvetkovic A et al (2010) Microbial metalloproteomes are largely uncharacterized. Nature 466(7307):779–782
Lothian A et al (2013) Metalloproteomics: principles, challenges and applications to neurodegeneration. Front Aging Neurosci 5:35
Yannone SM et al (2012) Metals in biology: defining metalloproteomes. Curr Opin Biotechnol 23(1):89–95
Fujita T et al (2004) Delta-catenin/NPRAP (neural plakophilin-related armadillo repeat protein) interacts with and activates sphingosine kinase 1. Biochem J 382(Pt 2):717–723
Leclercq TM et al (2008) Eukaryotic elongation factor 1A interacts with sphingosine kinase and directly enhances its catalytic activity. J Biol Chem 283(15):9606–9614
Sun J et al (2006) FHL2/SLIM3 decreases cardiomyocyte survival by inhibitory interaction with sphingosine kinase-1. Circ Res 99(5):468–476
Fukuda Y et al (2004) Identification of PECAM-1 association with sphingosine kinase 1 and its regulation by agonist-induced phosphorylation. Biochim Biophys Acta 1636(1):12–21
Lacana E et al (2002) Cloning and characterization of a protein kinase A anchoring protein (AKAP)-related protein that interacts with and regulates sphingosine kinase 1 activity. J Biol Chem 277(36):32947–32953
Maceyka M et al (2004) Aminoacylase 1 is a sphingosine kinase 1-interacting protein. FEBS Lett 568(1–3):30–34
Zebol JR et al (2009) The CCT/TRiC chaperonin is required for maturation of sphingosine kinase 1. Int J Biochem Cell Biol 41(4):822–827
Hayashi S et al (2002) Identification and characterization of RPK118, a novel sphingosine kinase-1-binding protein. J Biol Chem 277(36):33319–33324
Maceyka M et al (2008) Filamin A links sphingosine kinase 1 and sphingosine-1-phosphate receptor 1 at lamellipodia to orchestrate cell migration. Mol Cell Biol 28(18):5687–5697
Olivera A et al (1998) Purification and characterization of rat kidney sphingosine kinase. J Biol Chem 273(20):12576–12583
Jarman KE et al (2010) Translocation of sphingosine kinase 1 to the plasma membrane is mediated by calcium- and integrin-binding protein 1. J Biol Chem 285(1):483–492
Yamane D et al (2009) Inhibition of sphingosine kinase by bovine viral diarrhea virus NS3 is crucial for efficient viral replication and cytopathogenesis. J Biol Chem 284(20):13648–13659
Barr RK et al (2008) Deactivation of sphingosine kinase 1 by protein phosphatase 2A. J Biol Chem 283(50):34994–35002
Urtz N et al (2004) Early activation of sphingosine kinase in mast cells and recruitment to FcepsilonRI are mediated by its interaction with Lyn kinase. Mol Cell Biol 24(19):8765–8777
Olivera A et al (2006) IgE-dependent activation of sphingosine kinases 1 and 2 and secretion of sphingosine 1-phosphate requires Fyn kinase and contributes to mast cell responses. J Biol Chem 281(5):2515–2525
Xia P et al (2002) Sphingosine kinase interacts with TRAF2 and dissects tumor necrosis factor-alpha signaling. J Biol Chem 277(10):7996–8003
Gamble JR et al (2009) Sphingosine kinase-1 associates with integrin {alpha}V{beta}3 to mediate endothelial cell survival. Am J Pathol 175(5):2217–2225
Laviad EL et al (2012) Modulation of ceramide synthase activity via dimerization. J Biol Chem 287(25):21025–21033
Azuma N et al (1994) Stimulation of acid ceramidase activity by saposin D. Arch Biochem Biophys 311(2):354–357
Adam-Klages S et al (1996) FAN, a novel WD-repeat protein, couples the p55 TNF-receptor to neutral sphingomyelinase. Cell 86(6):937–947
Ahn KH et al (2013) Identification of heat shock protein 60 as a regulator of Neutral Sphingomyelinase 2 and its role in Dopamine uptake. PLoS One 8(6), e67216
Miteva YV, Budayeva HG, Cristea IM (2013) Proteomics-based methods for discovery, quantification, and validation of protein-protein interactions. Anal Chem 85(2):749–768
Paoletti AC et al (2006) Quantitative proteomic analysis of distinct mammalian Mediator complexes using normalized spectral abundance factors. Proc Natl Acad Sci U S A 103(50):18928–18933
Wang M et al (2012) PaxDb, a database of protein abundance averages across all three domains of life. Mol Cell Proteomics 11(8):492–500
Teo G et al (2014) SAINTexpress: improvements and additional features in significance analysis of INTeractome software. J Proteomics 100:37–43
Choi H et al (2011) SAINT: probabilistic scoring of affinity purification-mass spectrometry data. Nat Methods 8(1):70–73
Sowa ME et al (2009) Defining the human deubiquitinating enzyme interaction landscape. Cell 138(2):389–403
Rinner O et al (2007) An integrated mass spectrometric and computational framework for the analysis of protein interaction networks. Nat Biotechnol 25(3):345–352
Jager S et al (2011) Global landscape of HIV-human protein complexes. Nature 481(7381):365–370
Tackett AJ et al (2005) I-DIRT, a general method for distinguishing between specific and nonspecific protein interactions. J Proteome Res 4(5):1752–1756
Selbach M, Mann M (2006) Protein interaction screening by quantitative immunoprecipitation combined with knockdown (QUICK). Nat Methods 3(12):981–983
Mellacheruvu D et al (2013) The CRAPome: a contaminant repository for affinity purification-mass spectrometry data. Nat Methods 10(8):730–736
Wang X, Huang L (2008) Identifying dynamic interactors of protein complexes by quantitative mass spectrometry. Mol Cell Proteomics 7(1):46–57
Jurneczko E, Barran PE (2011) How useful is ion mobility mass spectrometry for structural biology? The relationship between protein crystal structures and their collision cross sections in the gas phase. Analyst 136(1):20–28
Lanucara F et al (2014) The power of ion mobility-mass spectrometry for structural characterization and the study of conformational dynamics. Nat Chem 6(4):281–294
Huang RY, Chen G (2014) Higher order structure characterization of protein therapeutics by hydrogen/deuterium exchange mass spectrometry. Anal Bioanal Chem 406(26):6541–6558
Jaswal SS (1834) Biological insights from hydrogen exchange mass spectrometry. Biochim Biophys Acta 6:1188–1201
Jorgensen TJ et al (2005) Intramolecular migration of amide hydrogens in protonated peptides upon collisional activation. J Am Chem Soc 127(8):2785–2793
Jorgensen TJ et al (2005) Collisional activation by MALDI tandem time-of-flight mass spectrometry induces intramolecular migration of amide hydrogens in protonated peptides. Mol Cell Proteomics 4(12):1910–1919
Zehl M et al (2008) Electron transfer dissociation facilitates the measurement of deuterium incorporation into selectively labeled peptides with single residue resolution. J Am Chem Soc 130(51):17453–17459
Rand KD et al (2008) Electron capture dissociation proceeds with a low degree of intramolecular migration of peptide amide hydrogens. J Am Chem Soc 130(4):1341–1349
Rappsilber J (2011) The beginning of a beautiful friendship: cross-linking/mass spectrometry and modelling of proteins and multi-protein complexes. J Struct Biol 173(3):530–540
Stengel F, Aebersold R, Robinson CV (2012) Joining forces: integrating proteomics and cross-linking with the mass spectrometry of intact complexes. Mol Cell Proteomics 11(3):R111.014027<>
Bruce JE (2012) In vivo protein complex topologies: sights through a cross-linking lens. Proteomics 12(10):1565–1575
Paramelle D et al (2013) Chemical cross-linkers for protein structure studies by mass spectrometry. Proteomics 13(3–4):438–456
Leitner A et al (2014) Chemical cross-linking/mass spectrometry targeting acidic residues in proteins and protein complexes. Proc Natl Acad Sci U S A 111(26):9455–9460
Fischer L, Chen ZA, Rappsilber J (2013) Quantitative cross-linking/mass spectrometry using isotope-labelled cross-linkers. J Proteomics 88:120–128
Hankins JL et al (2013) Ceramide 1-phosphate mediates endothelial cell invasion via the annexin a2-p11 heterotetrameric protein complex. J Biol Chem 288(27):19726–19738
Alvarez SE et al (2010) Sphingosine-1-phosphate is a missing cofactor for the E3 ubiquitin ligase TRAF2. Nature 465(7301):1084–1088
Heinrich M et al (1999) Cathepsin D targeted by acid sphingomyelinase-derived ceramide. EMBO J 18(19):5252–5263
Huwiler A et al (1996) Ceramide-binding and activation defines protein kinase c-Raf as a ceramide-activated protein kinase. Proc Natl Acad Sci U S A 93(14):6959–6963
Zhang Y et al (1997) Kinase suppressor of Ras is ceramide-activated protein kinase. Cell 89(1):63–72
Galadari S et al (1998) Purification and characterization of ceramide-activated protein phosphatases. Biochemistry 37(32):11232–11238
Lozano J et al (1994) Protein kinase C zeta isoform is critical for kappa B-dependent promoter activation by sphingomyelinase. J Biol Chem 269(30):19200–19202
Bourbon NA, Yun J, Kester M (2000) Ceramide directly activates protein kinase C zeta to regulate a stress-activated protein kinase signaling complex. J Biol Chem 275(45):35617–35623
Woodcock JM et al (2010) Sphingosine and FTY720 directly bind pro-survival 14-3-3 proteins to regulate their function. Cell Signal 22(9):1291–1299
Borch J, Roepstorff P, Moller-Jensen J (2011) Nanodisc-based co-immunoprecipitation for mass spectrometric identification of membrane-interacting proteins. Mol Cell Proteomics 10(7):O110.006775
Kota V, Szulc ZM, Hama H (2012) Identification of C(6)-ceramide-interacting proteins in D6P2T Schwannoma cells. Proteomics 12(13):2179–2184
Habrukowich C et al (2010) Sphingosine interaction with acidic leucine-rich nuclear phosphoprotein-32A (ANP32A) regulates PP2A activity and cyclooxygenase (COX)-2 expression in human endothelial cells. J Biol Chem 285(35):26825–26831
Stiban J, Tidhar R, Futerman AH (2010) Ceramide synthases: roles in cell physiology and signaling. Adv Exp Med Biol 688:60–71
Vunnam RR, Radin NS (1979) Short chain ceramides as substrates for glucocerebroside synthetase. Differences between liver and brain enzymes. Biochim Biophys Acta 573(1):73–82
Wijesinghe DS et al (2005) Substrate specificity of human ceramide kinase. J Lipid Res 46(12):2706–2716
Van Overloop H, Gijsbers S, Van Veldhoven PP (2006) Further characterization of mammalian ceramide kinase: substrate delivery and (stereo)specificity, tissue distribution, and subcellular localization studies. J Lipid Res 47(2):268–283
Senkal CE et al (2011) Alteration of ceramide synthase 6/C16-ceramide induces activating transcription factor 6-mediated endoplasmic reticulum (ER) stress and apoptosis via perturbation of cellular Ca2+ and ER/Golgi membrane network. J Biol Chem 286(49):42446–42458
Delon C et al (2004) Sphingosine kinase 1 is an intracellular effector of phosphatidic acid. J Biol Chem 279(43):44763–44774
Lee SJ et al (2014) Probing conformational change of intrinsically disordered alpha-synuclein to helical structures by distinctive regional interactions with lipid membranes. Anal Chem 86(3):1909–1916
Marcoux J et al (2013) Mass spectrometry reveals synergistic effects of nucleotides, lipids, and drugs binding to a multidrug resistance efflux pump. Proc Natl Acad Sci U S A 110(24):9704–9709
Eckford PD, Sharom FJ (2005) The reconstituted P-glycoprotein multidrug transporter is a flippase for glucosylceramide and other simple glycosphingolipids. Biochem J 389(Pt 2):517–526
Ciociola AA, Cohen LB, Kulkarni P (2014) How drugs are developed and approved by the FDA: current process and future directions. Am J Gastroenterol 109(5):620–623
Allende ML et al (2004) Mice deficient in sphingosine kinase 1 are rendered lymphopenic by FTY720. J Biol Chem 279(50):52487–52492
Bandhuvula P et al (2005) The immune modulator FTY720 inhibits sphingosine-1-phosphate lyase activity. J Biol Chem 280(40):33697–33700
Tonelli F et al (2010) FTY720 and (S)-FTY720 vinylphosphonate inhibit sphingosine kinase 1 and promote its proteasomal degradation in human pulmonary artery smooth muscle, breast cancer and androgen-independent prostate cancer cells. Cell Signal 22(10):1536–1542
Lahiri S et al (2009) Ceramide synthesis is modulated by the sphingosine analog FTY720 via a mixture of uncompetitive and noncompetitive inhibition in an Acyl-CoA chain length-dependent manner. J Biol Chem 284(24):16090–16098
Payne SG et al (2007) The immunosuppressant drug FTY720 inhibits cytosolic phospholipase A2 independently of sphingosine-1-phosphate receptors. Blood 109(3):1077–1085
Paugh SW et al (2006) Sphingosine and its analog, the immunosuppressant 2-amino-2-(2-[4-octylphenyl]ethyl)-1,3-propanediol, interact with the CB1 cannabinoid receptor. Mol Pharmacol 70(1):41–50
Cingolani F et al (2014) Inhibition of dihydroceramide desaturase activity by the sphingosine kinase inhibitor SKI II. J Lipid Res 55(8):1711–1720
Wang K et al (2012) Chemistry-based functional proteomics for drug target deconvolution. Expert Rev Proteomics 9(3):293–310
Liu Y, Guo M (2014) Chemical proteomic strategies for the discovery and development of anticancer drugs. Proteomics 14(4–5):399–411
Lukman S, Verma CS, Fuentes G (2014) Exploiting protein intrinsic flexibility in drug design. Adv Exp Med Biol 805:245–269
Huber W, Mueller F (2006) Biomolecular interaction analysis in drug discovery using surface plasmon resonance technology. Curr Pharm Des 12(31):3999–4021
Strickland EC et al (2013) Thermodynamic analysis of protein-ligand binding interactions in complex biological mixtures using the stability of proteins from rates of oxidation. Nat Protoc 8(1):148–161
Tran DT, Adhikari J, Fitzgerald MC (2014) SILAC-based strategy for proteome-wide thermodynamic analysis of protein-ligand binding interactions. Mol Cell Proteomics 13(7):1800–1813
West GM et al (2010) Quantitative proteomics approach for identifying protein-drug interactions in complex mixtures using protein stability measurements. Proc Natl Acad Sci U S A 107(20):9078–9082
Acknowledgments
This work was supported by the American Cancer Society and P01 CA171983. The authors would like to thank Kevin Fox for the creation of Fig. 1.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer International Publishing Switzerland
About this chapter
Cite this chapter
Stanley, B.A., Deering, T., Fox, T.E. (2015). Sphingoproteomics: Proteomic Strategies to Examine Sphingolipid Biology. In: Hannun, Y., Luberto, C., Mao, C., Obeid, L. (eds) Bioactive Sphingolipids in Cancer Biology and Therapy. Springer, Cham. https://doi.org/10.1007/978-3-319-20750-6_16
Download citation
DOI: https://doi.org/10.1007/978-3-319-20750-6_16
Publisher Name: Springer, Cham
Print ISBN: 978-3-319-20749-0
Online ISBN: 978-3-319-20750-6
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)