Highly efficient tandem affinity purification of trypanosome protein complexes based on a novel epitope combination.

ABSTRACT: Tandem affinity purification (TAP) allows for rapid and efficient purification of epitope-tagged protein complexes from crude extracts under native conditions. The method was established in yeast and has been successfully applied to other organisms, including mammals and trypanosomes. However, we found that the original method, which is based on the TAP tag, consisting of a duplicate protein A epitope, a tobacco etch virus protease cleavage site, and the calmodulin-binding peptide (CBP), did not yield enough recovery of transcription factor SNAPc (for small nuclear RNA-activating protein complex) from crude trypanosome extracts for protein identification. Specifically, the calmodulin affinity chromatography step proved to be inefficient. To overcome this problem, we replaced CBP by the pro

SUBMITTER: Schimanski B 

PROVIDER: S-EPMC1287860 | biostudies-literature | 2005 Nov

REPOSITORIES: biostudies-literature