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Li, Y., Tomko, R. J., & Hochstrasser, M. (2015). Proteasomes: Isolation and Activity Assays. Current Protocols In Cell Biology. Retrieved from http://purl.flvc.org/fsu/fd/FSU_pmch_26061243
In eukaryotes, damaged or unneeded proteins are typically degraded by the ubiquitin-proteasome system. In this system, the protein substrate is often first covalently modified with a chain of ubiquitin polypeptides. This chain serves as a signal for delivery to the 26S proteasome, a 2.5-MDa, ATP-dependent multisubunit protease complex. The proteasome consists of a barrel-shaped 20S core particle (CP) that is capped on one or both of its ends by a 19S regulatory particle (RP). The RP is responsible for recognizing the substrate, unfolding it, and translocating it into the CP for destruction. Here we describe simple, one-step purifications scheme for isolating the 26S proteasome and its 19S RP and 20S CP subcomplexes from the yeast Saccharomyces cerevisiae, as well as assays for measuring ubiquitin-dependent and ubiquitin-independent proteolytic activity in vitro.
Li, Y., Tomko, R. J., & Hochstrasser, M. (2015). Proteasomes: Isolation and Activity Assays. Current Protocols In Cell Biology. Retrieved from http://purl.flvc.org/fsu/fd/FSU_pmch_26061243
