Use of the novel technique of analytical ultracentrifugation with fluorescence detection system identifies a 77S monosomal translation complex.

ABSTRACT: A fundamental problem in proteomics is the identification of protein complexes and their components. We have used analytical ultracentrifugation with a fluorescence detection system (AU-FDS) to precisely and rapidly identify translation complexes in the yeast Saccharomyces cerevisiae. Following a one-step affinity purification of either poly(A)-binding protein (PAB1) or the large ribosomal subunit protein RPL25A in conjunction with GFP-tagged yeast proteins/RNAs, we have detected a 77S translation complex that contains the 80S ribosome, mRNA, and components of the closed-loop structure, eIF4E, eIF4G, and PAB1. This 77S structure, not readily observed previously, is consistent with the monosomal translation complex. The 77S complex abundance decreased with translational defects and followin

SUBMITTER: Wang X 

PROVIDER: S-EPMC3631355 | biostudies-literature | 2012 Sep

REPOSITORIES: biostudies-literature