Efficient biotinylation and single-step purification of tagged transcription factors in mammalian cells and transgenic mice.

ABSTRACT: Proteomic approaches require simple and efficient protein purification methodologies that are amenable to high throughput. Biotinylation is an attractive approach for protein complex purification due to the very high affinity of avidin/streptavidin for biotinylated templates. Here, we describe an approach for the single-step purification of transcription factor complex(es) based on specific in vivo biotinylation. We expressed the bacterial BirA biotin ligase in mammalian cells and demonstrated very efficient biotinylation of a hematopoietic transcription factor bearing a small (23-aa) artificial peptide tag. Biotinylation of the tagged transcription factor altered neither the factor's protein interactions or DNA binding properties in vivo nor its subnuclear distribution. Using this approac

SUBMITTER: de Boer E 

PROVIDER: S-EPMC164612 | biostudies-literature | 2003 Jun

REPOSITORIES: biostudies-literature