Large-scale detection of ubiquitination substrates using cell extracts and protein microarrays.

ABSTRACT: Identification of protein targets of post-translational modification is an important analytical problem in biology. Protein microarrays exposed to cellular extracts could offer a rapid and convenient means of identifying modified proteins, but this kind of biochemical assay, unlike DNA microarrays, depends on a faithful reconstruction of in vivo conditions. Over several years, concentrated cellular extracts have been developed, principally for cell cycle studies that reproduce very complex cellular states. We have used extracts that replicate the mitotic checkpoint and anaphase release to identify differentially regulated poyubiquitination. Protein microarrays were exposed to these complex extracts, and the polyubiquitinated products were detected by specific antibodies. We expected that a

SUBMITTER: Merbl Y 

PROVIDER: S-EPMC2633216 | biostudies-literature | 2009 Feb

REPOSITORIES: biostudies-literature