title: Analysis of chaperone-assisted ubiquitylation.
abstract: Molecular chaperones are traditionally viewed as cellular protein folding and assembly factors. However, in recent years it became more and more evident that certain chaperones, i.e., members of the 70-kDa heat shock protein family (Hsp70s), participate very actively in protein degradation and in this way significantly contribute to protein homeostasis. Degradation is often initiated through a close cooperation of Hsp70s with chaperone-associated ubiquitin ligases. This results in the ubiquitylation of chaperone-bound client proteins and triggers client sorting toward the proteasome or the autophagosome-lysosome system. Here, we describe the in vitro reconstitution of chaperone-assisted ubiquitylation, which allows analyzing molecular details of this important proteostasis mechanism.
authors.affiliation: Institut für Zellbiologie, Rheinische Friedrich-Wilhelms-Universität Bonn, Bonn, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Dreiseidler
authors.forename: Niko
authors.initials: N
authors.lastname: Dick
authors.forename: Jörg
authors.initials: J
authors.lastname: Höhfeld
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D051017:Apoptosis Regulatory Proteins; D001343:Autophagy; D002460:Cell Line; D050883:HSC70 Heat-Shock Proteins / Q000096:biosynthesis / Q000378:metabolism; D050956:HSP40 Heat-Shock Proteins / Q000096:biosynthesis / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D009036:Moths; D059748:Proteolysis; D019908:Proto-Oncogene Proteins c-raf / Q000378:metabolism; D051381:Rats; D044763:Ubiquitin-Conjugating Enzymes / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*; D054875:Ubiquitination / Q000502:physiology*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D051017:Apoptosis Regulatory Proteins; C414431:BAG3 protein, human; D050883:HSC70 Heat-Shock Proteins; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; C097309:UBE2D2 protein, human; D044763:Ubiquitin-Conjugating Enzymes; D044767:Ubiquitin-Protein Ligases; D019908:Proto-Oncogene Proteins c-raf