Semiquantitative proteomic analysis of the human spliceosome via a novel two-dimensional gel electrophoresis method.

ABSTRACT: More than 200 proteins associate with human spliceosomes, but little is known about their relative abundances in a given spliceosomal complex. Here we describe a novel two-dimensional (2D) electrophoresis method that allows separation of high-molecular-mass proteins without in-gel precipitation and thus without loss of protein. Using this system coupled with mass spectrometry, we identified 171 proteins altogether on 2D maps of stage-specific spliceosomal complexes. By staining with a fluorescent dye with a wide linear intensity range, we could quantitate and categorize proteins as present in high, moderate, or low abundance. Affinity-purified human B, B(act), and C complexes contained 69, 63, and 72 highly/moderately abundant proteins, respectively. The recruitment and release of spliceos

SUBMITTER: Agafonov DE 

PROVIDER: S-EPMC3133382 | biostudies-literature | 2011 Jul

REPOSITORIES: biostudies-literature