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Protein identification with N and C-terminal sequence tags in proteome projects
M R Wilkins1, E Gasteiger, L Tonella
1Central Clinical Chemistry Laboratory, Geneva University Hospital, 24 Rue Micheli-du-Crest, Geneva 14, 1211, Switzerland.
Journal of Molecular Biology
|June 20, 1998
Summary
Short protein sequence tags, like N-terminal and C-terminal tags, can identify proteins separated by 2D gel electrophoresis. This method, especially for prokaryotes, aids in proteome analysis and protein identification.
Area of Science:
- Proteomics
- Bioinformatics
- Genomics
Background:
- Increasing availability of genome sequences enables large-scale proteome studies.
- Two-dimensional (2D) gel electrophoresis is a common technique for protein separation.
- Accurate protein identification is crucial for understanding cellular functions.
Purpose of the Study:
- To investigate the utility of short N-terminal and C-terminal sequence tags for protein identification.
- To develop a computational tool for protein identification using sequence tags.
- To assess the applicability of this method across different species.
Main Methods:
- Analysis of theoretical N- and C-terminal sequences from SWISS-PROT database entries.
- Development of the TagIdent web-accessible program for protein identification.
- Experimental validation using 2D gel-purified Escherichia coli proteins.
Main Results:
- Sequence tags (4-6 amino acids) demonstrate high specificity for protein identification.
- TagIdent program successfully identified proteins using sequence tags, pI, and mass.
- Identification accuracy varied between prokaryotic and eukaryotic species.
Conclusions:
- Sequence tag-based protein identification is effective, particularly for prokaryotes with complete genome sequences.
- The TagIdent approach offers a valuable tool for proteomic research.
- Limitations exist for eukaryotic proteins due to sequencing challenges and incomplete genome data.