Summary
Proteomic studies of the free-living nematode Caenorhabditis elegans have recently received great attention because this animal model is a useful platform for the in vivo study of various biological problems relevant to human disease. In general, proteomic analysis is carried out in order to address a specific question with respect to differential changes in proteome expression under certain perturbed conditions. In this chapter, we focus on gel-based proteomic analysis of C. elegans subjected to two specific stress conditions during development: induction of the dauer state for whole body protein expression and a temperature shift for egg protein expression. Utilizing these differently perturbed C. elegans protein samples, two-dimensional electrophoresis and differential in-gel electrophoresis methods have led to the discovery of remarkable aspects of the worm’s biology. We also provide numerous details about the technical points and protocols necessary for successful experimentation.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
Notes
- 1.
This Method is useful for Preparation of large quantities of protein extracts from dauer larvae
References
Riddle, D., and Albert, P. (1997) C. elegans II (Riddle, D., Blumenthal, T., Meyer, B., and Priess, J., eds.), Cold Spring Harbor Laboratory Press, pp. 739–768
Paik, Y. K., Jeong, S. K., Lee, E. Y., Jeong, P. Y., and Shim, Y. H. (2006) C. elegans: an invaluable model organism for the proteomics studies of the cholesterol-mediated signaling pathway. Expert Rev Proteomics 3, 439–453
Cassada, R. C., and Russell, R. L. (1975) The dauer larva, a post-embryonic developmental variant of the nematode Caenorhabditis elegans. Dev Biol 46, 326–342
Horwitz, J. (1992) α-Crystallin can function as a molecular chaperone. Proc Natl Acad Sci U S A 89, 10449–10453
Veinger, L., Diamant, S., Buchner, J., and Goloubinoff, P. (1998) The small heat-shock protein IbpB from Escherichia coli stabilizes stress-denatured proteins for subsequent refolding by a multichaperone network. J Biol Chem 273, 11032–11037
Holt, S. J., and Riddle, D. L. (2003) SAGE surveys C. elegans carbohydrate metabolism: evidence for an anaerobic shift in the long-lived dauer larva. Mech Ageing Dev 124, 779–800
Wang, J., and Kim, S. K. (2003) Global analysis of dauer gene expression in Caenorhabditis elegans. Development 130, 1621–1634
Wadsworth, W. G., and Riddle, D. L. (1989) Developmental regulation of energy metabolism in Caenorhabditis elegans. Dev Biol 132, 167–173
Raz, A., Zhu, D. G., Hogan, V., Shah, N., Raz, T., Karkash, R., et al. (1990) Evidence for the role of 34-kDa galactoside-binding lectin in transformation and metastasis. Int J Cancer 46, 871–877
Perillo, N. L., Pace, K. E., Seilhamer, J. J., and Baum, L. G. (1995) Apoptosis of T cells mediated by galectin-1. Nature 378, 736–739
Alban, A., David, S. O., Bjorkesten, L., Andersson, C., Sloge, E., Lewis, S., et al. (2003) A novel experimental design for comparative two-dimensional gel analysis: Two-dimensional difference gel electrophoresis incorporating a pooled internal standard. Proteomics 3, 36–44
Brenner, S. (1974) The genetics of Caenorhabditis elegans. Genetics 77, 71–94
Choi, B. K., Shin, Y. K., Lee, E. Y., Jeong, P. Y., Shim, Y. H., Chitwood, D., and Paik, Y. K. (2008) Proteomic analysis of the sterol-mediated signaling pathways in C. elegans. Methods Mol Biol, Humana Press, 462 (11), 167–179
Cho, S. Y., Lee, E. Y., Kim, H. Y., Kang, M. J.,Lee, H. J., Kim, H., et al. (2008) Protein profiling of human plasma samples by two-dimensional electrophoresis. Methods Mol Biol, Humana Press, 428, 57–75
Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72, 248–254
Epstein, H. F., and Shakes, D. C. (1995) Caenorhabditis elegans: modern biological analysis of an organism. In: Cell Biology, Academic Press, London
Larsen, M. R., Sorensen, G. L., Fey, S. J., Larsen, P. M., and Roepstorff, P. (2001) Phosphoproteomics: evaluation of the use of enzymatic de-phosphorylation and differential mass spectrometric peptide mass mapping for site specific phosphorylation assignment in proteins separated by gel electrophoresis. Proteomics 1, 223–238
Acknowledgments
This study was supported by a grant from the Korean Health 21 R&D project, Ministry of Health & Welfare, Republic of Korea (A030003 to YKP), by the Technology Development Program of the Ministry of Agriculture and Forestry, Republic of Korea (606001–53–1-SB010 to YKP), and by a grant from the Basic Research Program that is supported by KOSEF (RO1–2005–000–11021–0 to YHS).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Jeong, PY., Na, K., Jeong, MJ., Chitwood, D., Shim, YH., Paik, YK. (2009). Proteomic Analysis of Caenorhabditis elegans . In: Tyther, R., Sheehan, D. (eds) Two-Dimensional Electrophoresis Protocols. Methods in Molecular Biology, vol 519. Humana Press. https://doi.org/10.1007/978-1-59745-281-6_10
Download citation
DOI: https://doi.org/10.1007/978-1-59745-281-6_10
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-58829-937-6
Online ISBN: 978-1-59745-281-6
eBook Packages: Springer Protocols