Mallory body filaments become insoluble after normal assembly into intermediate filaments

The deposition of 8-to-10-nm filaments into inclusion bodies is a fundamental cellular change that occurs in several degenerative processes of many tissues. However, little is known about the pathological filaments including whether the filaments assemble by the same mechanisms that govern the assembly of normal intermediate filaments. We have addressed this issue by studying the in vitro reassembly of the cytokeratin filaments that are deposited into experimental murine Mallory bodies (MBs) but have not yet become covalently crosslinked components of the MB. The reassembly process of both normal hepatocellular and MB-derived cytokeratins (CKs) was similar and characterized by a hierarchy of protofilament and protofibrils with a prominent axial periodicity of approximately 21 nm (normal hepatocellular CK, 20.7 +/- 2 nm; MB-derived CK, 20.1 +/- 2 nm). Purified MB-derived CK and normal hepatocellular CK comigrated in polyacrylamide gel electrophoresis indicating composition by similar CK isoforms. These results indicate that intermediate filaments formed from MB-derived CK are indistinguishable from filaments assembled from normal CK. On this basis, we conclude that the intermediate filaments that form inclusion bodies are not aberrantly assembled but become aggregated and post-translationally modified after their initial formation.