Browse > Article
http://dx.doi.org/10.5352/JLS.2005.15.3.427

Proteome Analysis of Drosophila melanogaster Used 2-DE and MALDI- TOF-MS  

Park Jeong-Won (Biohub Co., Ltd.)
Cha Jae-Young (Biohub Co., Ltd.)
Song Jae-Young (Department of Microbiology, College of Medicine, Gyeongsang National University)
Kim Hee-Kyu (Department of Applied Biology and Environmental Science, Gyeongsang National University)
Kim Beom-Kyu (Biohub Co., Ltd.)
Jeon Beong-Sam (Biohub Co., Ltd.)
Publication Information
Journal of Life Science / v.15, no.3, 2005 , pp. 427-433 More about this Journal
Abstract
With the completely discovery of the Drosophila genome sequence, the next great challenge is to extract its biological information by systematic expression and to perform functional analysis of the gene. Here we reported a proteome analysis of D. melanogaster with two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS). The cell extracts of D. melanogaster, $200{\mu}g$ were resolved to more than 400 silver-stained spots by 2-DE. The most abundant protein spots were ranged from 4.0-7.5 of pI and from 15-90 kDa of molecular weight. The excised spots were destained and in-gel digested by trypsin. The masses of the resulting peptide mixtures were measured by MALDI-TOF-MS. Identified proteins were compared with measured peptide mass and a dynamic peptide searching database which is accessible via the internet. The results revealed that identified proteins were produced by 59 genes derived from 65 protein spots.
Keywords
Drosophila melanogaster; Proteomics; Two-dimensional electrophoresis; MALDI-TOF-MS;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Ordway, R. W., J. Pallanck, and B. Ganetzky. 1994. Neurally expressed Drosophila encoding homologs for the NSF and SNAP secretory proteins. Proc. Natl. Acad. Sci. USA. 91, 5715-5719
2 Shevchenko, A, M. Wilm, O. Vorm, and M. Mann. 1996. Mass spectrometric sequencing of proteins from silverstained polyacrylamide gels. Anal. Chem. 68, 850-858   DOI   ScienceOn
3 Talluri, S., A Bhatt, and D. P. Smith. 1995. Identification of a Drosophila G protein a subunit (dGqa-3) expressed in chemosensory cells and central neurons. Proc. Natl. Acad. Sci. USA. 92, 11475-11479
4 Theurkauf, W. E., H. Baum, J. Bo, and P. C. Wensink. 1986. Tissue-specific and constitutive alpha-tubulin genes of Drosophila melanogaster code for structurally distinct proteins. Proc. Natl. Acad. Sci. USA. 83, 8477-8481
5 Vierstraete, E., P. Verleyen, G. Baggerman, W. D'Hertog, G. Van den Bergh, J. Arckens, A De Loof, and J. Schoofs. 2004. A proteomic approach for the analysis of instantly released wound and immune proteins in Drosophila rnelanogaster hemolymph. Proc. Natl. Acad. Sci. USA. 101, 470-475
6 Weekes, J., C. H. Wheeler, J. X. Yan, J. Weil, T. Eschenhagen, G. Scholtysik, and M. J. Dunn. 1999. Bovine dilated cardiomyopathy: proteomic analysis of an animal model of human dilated cardiomyopathy. Electrophoresis 20, 898-906   DOI   ScienceOn
7 Wilkins, M. R., C. Pasquali, R. D. Appel, K. au, O. Golaz, J. C. Sanchez, J. X. Yan, A A Gooley, G. Hughes, J. Humphery-Smith, K. J. Williams, and D. F. Hochstrasser. 1996. From proteins to proteomes: large scale protein identification by two-dimensional electrophoresis and amino acid analysis. Biotechnology 14, 61-65   DOI   ScienceOn
8 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   DOI   ScienceOn
9 Ericsson, C., Z. Petho, and H. Mehlin. 1997. An on-line two-dimensional polyacrylamide gel electrophoresis protein database of adults Drosophila melanogaster. Electrophoresis 18, 484-490   DOI   ScienceOn
10 Giot, I., J. S. Bader, C. Brouwer, et al. 2003. A protein interaction map of Drosophila melanogaster. Science 302, 1727-1736   DOI
11 Gorg, A., W. Postel, S. Gunther, J. Weser, J. R. Strahler, S. M. Hanash, J. Somerlat, and R. Kuick. 1988. Approach to stationary two-dimensional pattern: influence of focusing time and immobilinel carrier ampholytes concentrations. Electrophoresis 9, 37-46   DOI   ScienceOn
12 Heukeshoven, J. and R. Dernick. 1988. Improved silver staining procedure for fast staining in PhastSystem Development Unit. J. Staining of sodium dodecyl sulfate gels. Electrophoresis 9, 28-32   DOI   ScienceOn
13 http://www.proteinprospector.ucsf.edu
14 Laemmli, U. K. 1970. Oeavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685   DOI   ScienceOn
15 Bossie, C. A. and M. M. Sanders. 1993. A cDNA from Drosophila melanogaster encodes a lamin C-like intermediate filament protein. J. Cell Sci. 104, 1263-1272
16 Langen, H., C. Gray, D. Roder, J. F. Juranville, B. Takacs, and M. Fountoulakis. 1997. From genome to proteome: protein map of Haemophilus injluenzae. Electrophoresis 18, 1184-1192   DOI   ScienceOn
17 O'Connell, K. J. and J. T. Stults. 1997. Identification of mouse liver proteins on two-dimensional electrophoresis gels by matrix-assisted laser desorption/ionization mass spectrometry of in situ enzymatic digests. Electrophoresis 18, 349-359   DOI   ScienceOn
18 Messina, A, M. Neri, F. Perosa, C. Caggese, M. Marino, R. Caizzi, and V. De Pinto. 1996. Cloning and chromosomal localization of a cDNA encoding a mitochondrial porin from Drosophila melanogaster. FEBS Letter. 384, 9-13   DOI   ScienceOn
19 Adams, M. D., S. E. Celniker, R. A. Holt, et al. 2000. The genome sequence of Drosophila melanogaster. Science 287, 2185-2195   DOI
20 Baggerman G., A. Cerstiaens, A. De Loof, and J. Schoofs. 2002. Peptidornics of the larval Drosophila melanogaster central nervous system. J. BioI. Chem. 277, 40368-40374   DOI   ScienceOn
21 Quadroni, M. and P. James. 1999. Proteomics and automation. Electrophoresis 20, 664-677   DOI   ScienceOn
22 Gong, I., M. Purl, M. Unlu, M. Young, K. Robertson, S. Viswanathan, A. Krishnaswamy, S. R. Dowd, and J. S. Minden. 2004. Drosophila ventral furrow morphogenesis: a proteomic analysis. Development 131, 643-656   DOI   ScienceOn
23 Vierstraete, E., A Cerstiaens, G. Baggerman, G. Van den Bergh, A De Loof, and J. Schoofs. 2003. Proteomics in Drosophila melanogaster: first 2D database of larval hemolymph proteins. Biochem. Biophys. Res. Commun. 304, 831-838   DOI   ScienceOn