Identification of Gal4 activation domain-binding proteins in the 26S proteasome by periodate-triggered cross-linking
- 30 September 2005
- journal article
- research article
- Published by Royal Society of Chemistry (RSC) in Molecular BioSystems
- Vol. 1 (5-6), 366-372
- https://doi.org/10.1039/b510019d
Abstract
A common occurrence in biology is that a regulatory peptide, protein, or small molecule regulates the activity of a large multi-protein complex through direct interactions with a protein(s) in that complex. To characterize the direct receptor of the regulatory molecule, one would ideally like to study the native system. We report here that periodate-triggered cross-linking of catechol-containing regulatory factors, followed by two-dimensional electrophoresis and Western blotting, is an effective method for the characterization of regulatory factor–protein interactions in the context of large multi-protein complexes. We demonstrate the utility of this methodology by identifying the Rpt6/Sug1 and Rpt4/Sug2 proteins as the direct targets of transcriptional activation domains in the 26S proteasome.Keywords
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