An MBoC Favorite: Proteasomal proteomics: identification of nucleotide-sensitive proteasome-interacting proteins by mass spectrometric analysis of affinity-purified proteasomes
نویسنده
چکیده
The proteasome is a complex multifunctional machine that destroys proteins marked for ubiquitin-mediated proteolysis. In this paper, the authors employ an elegant approach to isolate and define yeast proteasomes and their suite of interacting proteins (Verma et al., 2000). This paper has something for everyone. The method the authors developed has now become the standard in the field for rapid proteasome purification. They identified and validated a new subunit of the proteasome. And their work gave a powerful glimpse into the role of the proteasome as a node of intracellular protein interactions, with dozens of proteasome-interacting proteins (PIPs) associating with core proteasome subunits in a nucleotide-dependent manner. The technological achievement and unique biological insight provided by this study justify its place as one of MBoC’s most-cited articles.
منابع مشابه
Proteasomal proteomics: identification of nucleotide-sensitive proteasome-interacting proteins by mass spectrometric analysis of affinity-purified proteasomes.
Ubiquitin-dependent proteolysis is catalyzed by the 26S proteasome, a dynamic complex of 32 different proteins whose mode of assembly and mechanism of action are poorly understood, in part due to the difficulties encountered in purifying the intact complex. Here we describe a one-step affinity method for purifying intact 26S proteasomes, 19S regulatory caps, and 20S core particles from budding ...
متن کاملAffinity purification strategy to capture human endogenous proteasome complexes diversity and to identify proteasome-interacting proteins.
An affinity purification strategy was developed to characterize human proteasome complexes diversity as well as endogenous proteasome-interacting proteins (PIPs). This single step procedure, initially used for 20 S proteasome purification, was adapted to purify all existing physiological proteasome complexes associated to their various regulatory complexes and to their interacting partners. The...
متن کاملProteomic analysis of affinity-purified extracellular proteasomes reveals exclusively 20S complexes
Proteasome-mediated proteolysis is important for many basic cellular processes. In addition to their functions in the cell, proteasomes have been found in physiological fluids of both healthy and diseased humans including cancer patients. Higher levels of these proteasomes are associated with higher cancer burden and stage. The etiology and functions of these proteasomes, referred to as circula...
متن کاملCharacterization of the Brain 26S Proteasome and its Interacting Proteins
Proteasome-mediated proteolysis is important for synaptic plasticity, neuronal development, protein quality control, and many other processes in neurons. To define proteasome composition in brain, we affinity purified 26S proteasomes from cytosolic and synaptic compartments of the rat cortex. Using tandem mass spectrometry, we identified the standard 26S subunits and a set of 28 proteasome-inte...
متن کاملCharacterization of the Brain Proteasome and Its Interacting Proteins and Their Regulation by Neuronal Activity
Despite the importance of proteasome-mediated proteolysis in synaptic plasticity, protein quality control, and cell regulation, little is known about proteasome composition and regulation in the brain. This thesis represents the first detailed study of mammalian brain proteasomes. Using a new affinity purification method, 26S proteasomes were isolated from the cytosolic and the synaptic compart...
متن کامل