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 (Schägger, 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.
Athlan
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Department of Biochemistry, McGill University, 3655 Drummond Street, Montreal, Quebec H3G 1Y6, Canada.
Mushynski
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D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism; D002478:Cells, Cultured; D019281:Dimerization; D015180:Electrophoresis, Gel, Two-Dimensional; D007381:Intermediate Filament Proteins; D016900:Neurofilament Proteins / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D051381:Rats
D002352:Carrier Proteins; D007381:Intermediate Filament Proteins; D016900:Neurofilament Proteins; C046643:alpha-internexin; C044882:neurofilament protein L; C051774:neurofilament protein H; C061195:neurofilament protein M