抄録
Chemical cross-linking combined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) is a simple way to analyze oligomerization and complex formation of proteins. However, cross-linked proteins with a large molecular mass, for example with molecular weight greater than ∼400 kDa, show slow mobility and tend to stack in loading area with poor resolution. Agarose gel electrophoresis (AGE) may be more suitable, due to a large pore size of the agarose, to analyze proteins as well as DNA fragments, especially for large macromolecules and their complexes. Here, we expressed and purified recombinant ClpB chaperone protein (homologue of eukaryotic HSP104) derived from moderate halophile and analyzed the purified protein by SDS-PAGE and SDS-AGE. Chemical cross-linking and SDS-AGE analysis clearly demonstrated hexameric assembly of ClpB subunit protein. Heat-denaturation profile of the ClpB with native-AGE analysis done without SDS suggested that ClpB protein was denatured at around 50 °C.
