A two-dimensional electrophoresis reference map of the earthworm Eisenia fetida

Abstract

The molecular mechanisms underlying innate immunity in the earthworm E. fetida remain unclear. For the recognition of innate immunity in the earthworm E. fetida, a detailed knowledge of this proteome is a prerequisite. The absence of a high-resolution E. fetida proteome map prompted us to determine E. fetida protein spots that can be visualised on 2-D protein gels. In this study, we present a preliminary description of the whole earthworm E. fetida proteome. A highly detailed two-dimensional gel electrophoresis (2-DE) map of the E. fetida proteome was established and approximately 1500 protein spots were detected from the earthworm sample when applying a 500 μg protein 2-DE in the pH range 3.0 - 10.0. We present a 2-DE proteome map of E. fetida, identifying 76 different proteins by matrix-assisted laser desorption/ionization-tandem time of flight mass spectrometry (MALDI-TOF/ TOF-MS) analysis. These identified proteins, including heat shock protein 90 (Hsp90), chaperonine protein HSP60, caspase-8, fibrinolytic protease 0, gelsolin-like protein, lombricine kinase, coelomic cytolytic factor1 (CCF 1), manganous superoxide dismutase (MnSOD), triosephosphate isomerase, extracellular globin-4, lysenin, and intermediate filament protein, glyceraldehyde-3- phosphate dehydrogenase, et al., are involved in several processes, including transcripttion, translation, the tricarboxylic acid cycle, the cellular amino acid metabolic process, protein amino acid phosphorylation, glycolysis, and the glucose metabolic process. These 2-DE data will enhance future comparisons of immunity, toxicology, biotic processes and other challenges, thereby allowing for further study of the molecular mechanisms in response to environmental stressors, and it will be useful to investigate environmental proteomics in invertebrate earthworms.

Share and Cite:

Wang, X. , Zhang, Y. and Sun, Z. (2012) A two-dimensional electrophoresis reference map of the earthworm Eisenia fetida. Natural Science, 4, 869-880. doi: 10.4236/ns.2012.411115.

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1] Edwards, C.A. (2004) Earthworm ecology. 2nd Edition, CRC Press, Boca Raton.
[2] Pirooznia, M., Gong, P., Guan, X., Inouye, L.S., Yang, K., Perkins, E.J. and Deng, Y.P. (2009) Cloning, analysis and functional annotation of expressed sequence tags from the earthworm Eisenia fetida. BMC Bioinformatics, 8, 1-16. Hdoi:10.1186/1471-2105-8-S7-S7
[3] Bundy, J.G., Sidhu, J.K., Rana, F., Spurgeon, D.J., Svendsen, C.S., Wren, J.F. Stürzenbaum, S.R., Morgan, A. J. and Kille, P. (2008) “Systems toxicology” approach identifies coordinated metabolic responses to copper in a terrestrial non-model invertebrate, the earthworm Lumbricus rubellus. BMC Biology, 6, 25. Hdoi:10.1186/1741-7007-6-25
[4] Amelina, H., Apraiz, I., Sun, W. and Cristobal, S. (2007) Proteomics-based method for the assessment of marine pollution using liquid chromatography coupled with twodimensional electrophoresis. Journal of Proteome Research, 6, 2094-2104. Hdoi:10.1021/pr060689s
[5] Dowling, V.A. and Sheehan, D. (2006) Proteomics as a route to identification of toxicity targets in environmental toxicology. Proteomics, 6, 5597-5604. Hdoi:10.1002/pmic.200600274
[6] Lee, S.E., Yoo, D.H., Son, J. and Cho, K. (2006) Proteomic evaluation of cadmium toxicity on the midge Chironomus riparius Meigen larvae. Proteomics, 6, 945- 957. Hdoi:10.1002/pmic.200401349
[7] Knigge, T., Monsinjon, T. and Andersen, O.K. (2004) Surface-enhanced laser desorption/ionization-time of flightmass spectrometry approach to biomarker discovery in blue mussels [Mytilus edulis] exposed to polyaromatic hydrocarbons and heavy metals under field conditions. Proteomics, 4, 2722-2727. Hdoi:10.1002/pmic.200300828
[8] Owen, J., Hedley, B.A., Svendsen, C., Wren, J., Jonker, M. J., Hankard, P.K., Lister, L.J., Stürzenbaum, S.R., Morgan, A.J., Spurgeon, D.J., Blaxter, M.L. and Kille, P. (2008) Transcriptome profiling of developmental and xenobiotic responses in a keystone soil animal, the oligochaete annelid Lumbricus rubellus. BMC Genomics, 9, 226. Hdoi:10.1186/1471-2164-9-266
[9] Gong, P., Guan, X., Inouye, L.S., Deng, Y., Pirooznia, M. and Perkins, E.J. (2008) Transcriptomic analysis of RDX and TNT interactive sublethal effects in the earthworm Eisenia fetida. BMC Genomics, 9, S15. Hdoi:10.1186/1471-2164-9-S1-S15
[10] Nabby-Hansen, S., Waterfield, M.D. and Cramer, R. (2001) Proteomics-post-genomic cartography to understand gene function. Trends in Pharmacological Sciences, 22, 376- 384. Hdoi:10.1016/S0165-6147(00)01663-1
[11] Kuperman, R.G., Checkai, R.T., Ruth, L.M., Henry, T., Simini, M., Kimmel, D.G., Phillips, C.T. and Bradley, B.P. (2003) A proteome-based assessment of the earthworm Eisenia fetida: Response to chemical warfare agents in a sandy loam soil. Pedobiologia, 47, 617-621. Hdoi:10.1016/S0031-4056(04)70245-0
[12] LaCourse, E.J., Hernandez-Viadel, M., Jefferies, J.R., Svendsen, C., Spurgeon, D.J., Barrett, J., Morgan, A.J., Kille, P. and Brophy, P.M. (2009) Glutathione transferase [GST] as a candidate molecular-based biomarker for soil toxin exposure in the earthworm Lumbricus rubellus. Environmental Pollution, 157, 2459-2469. Hdoi:10.1016/j.envpol.2009.03.015H
[13] Wang, X., Chang, L., Wang, G.C., Sun, Z.J., Ma, H.B., Sun, Q. and Li, J. (2010) Protein extraction from the earthworm Eisenia fetida for 2-DE. Proteomics, 10, 1095- 1099. Hdoi:10.1002/pmic.200900488
[14] Wang, X., Chang, L., Sun, Z.J., Zhang, Y.F. and Yao, L. (2010) Analysis of earthworm Eisenia fetida proteomes during cadmium exposure: An ecotoxicoproteomics approach. Proteomics, 10, 4476-4490. Hdoi:10.1002/pmic.201000209
[15] Wang, X., Chang, L., Sun, Z.J. and Zhang, Y.F. (2010) Comparative proteomic analysis of differentially expressed proteins in the earthworm Eisenia fetida during Escherichia coli O157:H7 stress. Journal of Proteome Research, 9, 6547-6560. Hdoi:10.1021/pr1007398
[16] Hernández, R., Nombela, C., Diez-Orejas, R. and Gil, C. (2004) Two-dimensional reference map of Candida albicans hyphal forms. Proteomics, 4, 374-382. Hdoi:10.1002/pmic.200300608
[17] OECD (2004) OECD (Organization for Economic Cooperation and Development) guideline for testing chemicals. Section 2: Effects on biotic systems. Earthworm, Acute Toxicity Tests, Paris.
[18] Carpentier, S.C., Witters, E., Laukens, K., Deckers, P., Swennen, R. and Panis, B. (2005) Preparation of protein extracts from recalcitrant plant tissues: An evaluation of different methods for two-dimensional gel electrophoresis analysis. Proteomics, 5, 2497-2507. Hdoi:10.1002/pmsic.200401222
[19] Lim, H., Eng, J., Yates, J.R., Tollaksen, S.L., Giometti, C. S., Holden, J.F., Adams, M.W.W., Reich, C.I., Olsen, G.J. and Hays, L.G. (2003) Identification of 2D-gel proteins: A comparison of MALDI/TOF peptide mass mapping to mu LC-ESI tandem mass spectrometry. Journal of the American Society for Mass Spectrometry, 14, 957-970. Hdoi:10.1016/S1044-0305(03)00144-2
[20] Huang, F., Fulda, S., Hagemann, M. and Norling, B. (2006) Proteomic screening of salt-stress-induced changes in plasma membranes of Synechocystis sp. strain PCC 6803. Proteomics, 6, 910-920. Hdoi:10.1002/pmic.200500114
[21] Sarry, J.E., Kuhn, L., Ducruix, C., Lafaye, A., Junot, C., Hugouvieux, V., Jourdain, A., Bastien, O., Fievet, J.B., Vailhen, D., Amekraz, B., Moulin, C., Ezan, E., Garin, J. and Bourguignon, J. (2006) The early responses of Arabidopsis thaliana cells to cadmium exposure explored by protein and metabolite profiling analyses. Proteomics, 6, 2180-2198. Hdoi:10.1002/pmic.200500543
[22] Nishi, H., Inagi, R., Kato, H., Tanemoto, M., Kojima, I., Son, D., Fujita, T. and Nangaku, M. (2008) Hemoglobin is expressed by mesangial cells and reduces oxidant stress. Journal of the American Society of Nephrology, 19, 1500- 1508. Hdoi:10.1681/ASN.2007101085H
[23] McGregor, E., Kempster, L., Wait, R., Gosling, M., Dunn, M.J. and Powell, J.T. (2004) F-actin capping [CapZ] and other contractile saphenous vein smooth muscle proteins are altered by hemodynamic stress. Molecular & Cellular Proteomics, 3, 115-124. Hdoi:10.1074/mcp.M300046-MCP2004
[24] Sekizawa, Y., Kubo, T. and Kobayashi, H. (1997) Molecular cloning of cDNA for lysenin, a novel protein in the earthworm Eisenia foetida that causes contraction of rat vascular smooth muscle. Gene, 191, 97-102. Hdoi:10.1016/S0378-1119(97)00047-4
[25] Yamaji, A., Sekizawa, Y., Emoto, K., Sakuraba, H., Inoue, K., Kobayashi, H. and Umeda, M. (1998) Lysenin, a novel sphingomyelin-specific binding protein. Journal of Biological Chemistry, 273, 5300-5306. Hdoi:10.1074/jbc.273.9.5300
[26] K?hlerová, P., Beschin, A., ?ilerová, M., De Baetselier, P. and Bilej, M. (2004) Effect of experimental microbial challenge on the expression of defense molecules in Eisenia foetida earthworm. Developmental & Comparative Immunology, 28, 701-711. Hdoi:10.1016/j.dci.2004.01.001
[27] Zhang, X.M., Luan, W., Jin, S.J. and Xiang, J. (2008) A novel tumor necrosis factor ligand superfamily member [CsTL] from Ciona savignyi: molecular identification and expression analysis. Developmental & Comparative Immunology, 32, 1362-1373. Hdoi:10.1016/j.dci.2008.05.009
[28] Lynch, J.A., Brent, A.E., Leaf, D.S., Pultz, M.A. and Desplan, C. (2006) Localized maternal orthodenticle patterns anterior and posterior in the long germ wasp Nasonia. Nature, 39, 728-732. Hdoi:10.1038/nature04445
[29] T?rr?nen, R., Korkalainen, M. and K?renlampi, S.O. (1992) Induction of class 3 aldehyde dehydrogenase in the mouse hepatoma cell line Hepa-1 by various chemicals. Chemico-Biological Interactions, 83, 107-119. Hdoi:10.1016/0009-2797(92)90040-R
[30] McKim, J.M., Choudhuri, S. and Klaassen, C.D. (1992) In vitro degradation of apo-, zinc-, and cadmium-metallothionein by cathepsins B, C, and D. Toxicology and Applied Pharmacology, 116, 117-124. Hdoi:10.1016/0041-008X(92)90152-I
[31] Martens, S. and Howard, J. (2006) The interferon-inducible GTPases. Annual Review of Cell and Developmental Biology, 22, 559-589. Hdoi:10.1146/annurev.cellbio.22.010305.104619
[32] Djabali, K., de Nechaud, B., Landon, F. and Portier, M.M. (1997) AlphaB-crystallin interacts with intermediate filaments in response to stress. Journal of Cell Science, 110, 2759-2769.
[33] Ellington, W.R. (2001) Evolution and physiological roles of phosphagen systems. Annual Review of Physiology, 63, 289-325.Hdoi:10.1146/annurev.physiol.63.1.289H
[34] Park, Y., Ryu, E., Kim, H., Jeong, J., Kim, J., Shim, J., Jeon, S., Jo, Y., Kim, W. and Min, B. (1999) Characterization of antithrombotic activity of lumbrokinase-immobilized polyurethane valves in the total artificial heart. Artificial Organs, 23, 210-214. Hdoi:10.1046/j.1525-1594.1999.06013.x
[35] Boldin, M.P., Goncharov, T.M., Goltsev, Y.V. and Wallach, D. (1996) Involvement of MACH, a novel MORT1/ FADD-interacting protease, in Fas/APO-1- and TNF re- ceptor-induced cell death. Cell, 85, 803-815. Hdoi:10.1016/S0092-8674(00)81265-9H
[36] Hengartner, M.O. (2000) The biochemistry of apoptosis. Nature, 407, 770-776. Hdoi:10.1038/35037710
[37] Salvesen, G.S. and Dixit, V.M. (1997) Caspases: Intracel- lular signaling by proteolysis. Cell, 91, 443-446. Hdoi:10.1016/S0092-8674(00)80430-4
[38] Ram, R.J., VerBerkmoes, N.C., Thelen, M.P., Tyson, G. W., Baker, B.J., Blake II, R.C., Shah, M., Hettich, R.L. and Banfield, J.F. (2005) Community proteomics of a natural microbial biofilm. Science, 308, 1915-1920. Hdoi:10.1126/science. 1109070
[39] Brown, P.J., Long, S.M., Spurgeon, D.J., Svendsen, C. and Hankard, P.K. (2004) Toxicological and biochemical responses of the earthworm Lumbricus rubellus to pyrene, a non-carcinogenic polycyclic aromatic hydrocarbon. Che- mosphere, 57, 1675-1681.
[40] Pandey, P., Saleh, A., Nakazawa, A., Kumar, S., Srini- vasula, S.M., Kumar, V., Weichselbaum, R., Nalin, C., Alnemri, E.S., Donald, K. and Kharbanda, S. (2000) Nega- tive regulation of cytochrome c-mediated oligomerization of Apaf-1 and activation of procaspase-9 by heat shock protein 90. The EMBO Journal, 19, 4310-4322. Hdoi:10.1093/emboj/19.16.4310
[41] Zhao, R., Davey, M., Hsu, Y.C., Kaplanek, P., Tong, A., Parsons, A.B., Krogan, N., Cagney, G., Mai, D., Green- blatt, J., Boone, C., Emili, A. and Houry, W.A. (2005) Navigating the chaperone network: An integrative map of physical and genetic interactions mediated by the Hsp90 chaperone. Cell, 120, 715-727. Hdoi:10.1016/j.cell.2004.12.024
[42] Cho, J.H., Park, C.B., Yoon, Y.G. and Kim, S.C. (1998) Lumbricin I, a novel proline-rich antimicrobial peptide from the earthworm: Purification, cDNA cloning and mo- lecular characterization. Biochimica et Biophysica Acta, 1408, 67-76. Hdoi:10.1016/S0925-4439(98)00058-1
[43] Kim, H.R., Han, R.X., Yoon, J.T., Park, C.S. and Jin, D.I. (2010) A two-dimensional electrophoresis reference map for the bovine placenta during late pregnancy. Proteomics, 10, 564-573. Hdoi:10.1002/pmic.200900508

Copyright © 2024 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.