Affinity purification strategy to capture human endogenous proteasome complexes diversity and to identify proteasome-interacting proteins.

ABSTRACT: 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 method was applied to the purification of proteasome complexes and their PIPs from human erythrocytes but can be used to purify proteasomes from any human sample as starting material. The benefit of in vivo formaldehyde cross-linking as a stabilizer of protein-protein interactions was studied by comparing the status of purified proteasomes and the identified proteins in both protocols (with or witho

SUBMITTER: Bousquet-Dubouch MP 

PROVIDER: S-EPMC2689779 | biostudies-literature | 2009 May

REPOSITORIES: biostudies-literature