title: Characterization of the proteasome interaction network using a QTAX-based tag-team strategy and protein interaction network analysis.
abstract: Quantitative analysis of tandem-affinity purified cross-linked (x) protein complexes (QTAX) is a powerful technique for the identification of protein interactions, including weak and/or transient components. Here, we apply a QTAX-based tag-team mass spectrometry strategy coupled with protein network analysis to acquire a comprehensive and detailed assessment of the protein interaction network of the yeast 26S proteasome. We have determined that the proteasome network is composed of at least 471 proteins, significantly more than the total number of proteins identified by previous reports using proteasome subunits as baits. Validation of the selected proteasome-interacting proteins by reverse copurification and immunoblotting experiments with and without cross-linking, further demonstrates the power of the QTAX strategy for capturing protein interactions of all natures. In addition, >80% of the identified interactions have been confirmed by existing data using protein network analysis. Moreover, evidence obtained through network analysis links the proteasome to protein complexes associated with diverse cellular functions. This work presents the most complete analysis of the proteasome interaction network to date, providing an inclusive set of physical interaction data consistent with physiological roles for the proteasome that have been suggested primarily through genetic analyses. Moreover, the methodology described here is a general proteomic tool for the comprehensive study of protein interaction networks.
authors.affiliation: Departments of Physiology and Biophysics and Developmental and Cell Biology, University of California, Irvine, CA 92697, USA.
authors.forename: Cortnie
authors.initials: C
authors.lastname: Guerrero
authors.forename: Tijana
authors.initials: T
authors.lastname: Milenkovic
authors.forename: Natasa
authors.initials: N
authors.lastname: Przulj
authors.forename: Peter
authors.initials: P
authors.lastname: Kaiser
authors.forename: Lan
authors.initials: L
authors.lastname: Huang