Heterodimeric associations between neuronal intermediate filament proteins.
Formation of protein dimers involving alpha-internexin, peripherin, and the neurofilament (NF) proteins NFH, NFM, and NFL was investigated by partial renaturation of various combinations of individually purified subunits in buffered 2 M urea. Oligomers that were formed were resolved by "blue" native electrophoresis (Schaegger, H., Cramer, W. A., and von ... Jagow, G. (1994) Anal. Biochem. 217, 220-230) modified to include urea in the polyacrylamide gels. Combining this method with Western blot analysis, disulfide cross-linking, and SDS-polyacrylamide gel electrophoresis in the second dimension showed that NFL readily forms significant amounts of heterodimer with NFH, NFM, alpha-internexin, or peripherin in the presence of 2 M urea. alpha-Internexin and peripherin also formed heterodimers with NFH or NFM under these conditions. The modified version of blue native gel electrophoresis described here may be useful in monitoring the impact of post-translational modifications and mutations on the dimerization of intermediate filament proteins.
Mesh Terms:
Animals, Carrier Proteins, Cells, Cultured, Dimerization, Electrophoresis, Gel, Two-Dimensional, Intermediate Filament Proteins, Neurofilament Proteins, Protein Binding, Protein Conformation, Rats
Animals, Carrier Proteins, Cells, Cultured, Dimerization, Electrophoresis, Gel, Two-Dimensional, Intermediate Filament Proteins, Neurofilament Proteins, Protein Binding, Protein Conformation, Rats
J. Biol. Chem.
Date: Dec. 05, 1997
PubMed ID: 9388258
View in: Pubmed Google Scholar
Download Curated Data For This Publication
5339
Switch View:
- Interactions 1