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The rat liver epithelial (RLE) cell nuclear protein database
P J Wirth1, L D Luo, T Benjamin
1Biopolymer Chemistry Section, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
Electrophoresis
|November 1, 1993
Summary
The expanded rat liver epithelial (RLE) nuclear protein database now details over 2000 polypeptides, aiding research into cellular transformation and growth. This comprehensive resource facilitates identification of key regulatory proteins.
Area of Science:
- Proteomics
- Cell Biology
- Biochemistry
Background:
- The rat liver epithelial (RLE) cellular protein database provides a foundation for studying protein expression.
- Previous work established a master database for RLE cellular proteins.
Purpose of the Study:
- To expand the RLE nuclear protein database with detailed proteomic and quantitative information.
- To characterize nucleoplasmic and particulate-associated RLE nuclear polypeptides.
- To facilitate the identification of transformation and growth-related proteins.
Main Methods:
- Utilized the Elsie 5 gel analysis system for database development.
- Incorporated [35S]methionine and [32P]orthophosphate labeling for polypeptide analysis.
- Implemented direct microsequencing from two-dimensional (2-D) blotted membranes for polypeptide identification.
Main Results:
- Detailed information on 1100 nucleoplasmic and 850 particulate-associated [35S]methionine labeled RLE nuclear polypeptides.
- Detailed information on 215 nucleoplasmic and 269 particulate-associated [32P]orthophosphate labeled RLE nuclear polypeptides.
- Database includes qualitative and quantitative annotations: identification number, molecular weight, pI, quantitation, spot shape, subcellular location, and transformation/growth characteristics.
Conclusions:
- The RLE nuclear protein database is a continually updated resource.
- Direct microsequencing offers rapid polypeptide identification without specific antibodies.
- Findings are applicable to other rodent and human 2-D protein databases, aiding research in cell transformation and growth regulation.