- - Site under construction. This page was last modified on 7 July 1997 - -
Dr. Mike Cahill
Stefanie Weiß
The implementation of new technologies and the explosion of information in the sequence data bases are changing the way that knowledge is perceived in the biological sciences. An example is provided by recent advances in protein two dimensional polyacrylamide gel electrophoresis (2D-PAGE) and protein sequencing technology. 2D-PAGE is now the system of choice for identifying molecules which are differentially represented in any two or more cell types, being well suited to the examination of intracellular processes involved in signal transduction, programmed cell death, differentiation, or disease progression. This emerging field has been given the acronym PROTEOME analysis (PROTEins within a genOME). The PROTEOME methodolgy may eventually reveal the identitiy and regulation of most proteins expressed within a particular cell-type (Wilkins et al., 1996; Pennington et al., 1997).
2D-PAGE involves separation of proteins in the first dimension by differential isoelectric points, and in the second dimension by molecular weight. This results in 2-D protein spots of near absolute biochemical purity, which facilitates subsequent sensitive analyses. Proteins of interest are isolated from the gel, and partial sequence information is gathered by either Edman sequencing coupled with amino acid analysis, or by Mass Spectrometry. This partial information is used to identify the protein by computer alignment with existing sequence data bases which are being generated by the genome projects (Lamond and Mann, 1997).
An intrinsic strength of 2D-PAGE as opposed to other methods is that it can reveal molecules which undergo post translational modifications, such as phosphorylation, geranylation, glycosylation, proteolytic cleavage, translational induction, alternative splicing, or any modification that affects molecular weight or isoelectric point. This developing technology essentially corresponds to a differential display on the protein level and looks poised to replace the nucleic acid-based method. Proteome analysis is an emerging and exciting new field which promises to significantly shift our horizons in the understanding of cell biology and disease processes.
At time of writing there are positions available for students (at any level) interested in entering this stimulating new field. If working in the 2D-electrophoresis group and the prospect of living in the beautiful city of Tuebingen interest you, then contact Dr. Mike Cahill for details.
Lamond, A.I. and Mann, M. (1997) Cell Biology and the genome projects- a concerted strategy for characterising multiprotein complexes using mass spectrometry. Trends Biochem. Sci. 7: 139142.
Pennington, S.R., Wilkins, M.R., Hochstrasse, D.F., and Dunn, M.J. (1997) Proteome analysis: from protein characterisation to biological function. Trends Cell Biol. 7: 168-173.
Wilkins, M.R., Sanchez, J.-C., Williams, K.L., Hochstrasse, D.F. (1996) Current challenges and future applications for protein maps and post-translational vector maps in proteome projects. Electrophoresis 17: 830-838.
